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1.
Curr Genet ; 70(1): 6, 2024 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-38733432

RESUMO

The gene products of PRS1-PRS5 in Saccharomyces cerevisiae are responsible for the production of PRPP (5-phospho-D-ribosyl-α-1-pyrophosphate). However, it has been demonstrated that they are also involved in the cell wall integrity (CWI) signalling pathway as shown by protein-protein interactions (PPIs) with, for example Slt2, the MAP kinase of the CWI pathway. The following databases: SGD, BioGRID and Hit Predict, which collate PPIs from various research papers, have been scrutinized for evidence of PPIs between Prs1-Prs5 and components of the CWI pathway. The level of certainty in PPIs was verified by interaction scores available in the Hit Predict database revealing that well-documented interactions correspond with higher interaction scores and can be graded as high confidence interactions based on a score > 0.28, an annotation score ≥ 0.5 and a method-based high confidence score level of ≥ 0.485. Each of the Prs1-Prs5 polypeptides shows some degree of interaction with the CWI pathway. However, Prs5 has a vital role in the expression of FKS2 and Rlm1, previously only documented by reporter assay studies. This report emphasizes the importance of investigating interactions using more than one approach since every method has its limitations and the use of different methods, as described herein, provides complementary experimental and statistical data, thereby corroborating PPIs. Since the experimental data described so far are consistent with a link between PRPP synthetase and the CWI pathway, our aim was to demonstrate that these data are also supported by high-throughput bioinformatic analyses promoting our hypothesis that two of the five PRS-encoding genes contain information required for the maintenance of CWI by combining data from our targeted approach with relevant, unbiased data from high-throughput analyses.


Assuntos
Parede Celular , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Parede Celular/metabolismo , Parede Celular/genética , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Transdução de Sinais , Mapas de Interação de Proteínas , Mapeamento de Interação de Proteínas
2.
Int J Mol Sci ; 25(9)2024 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-38732136

RESUMO

In the context of sustainable agriculture and biomaterial development, understanding and enhancing plant secondary cell wall formation are crucial for improving crop fiber quality and biomass conversion efficiency. This is especially critical for economically important crops like upland cotton (Gossypium hirsutum L.), for which fiber quality and its processing properties are essential. Through comprehensive genome-wide screening and analysis of expression patterns, we identified a particularly high expression of an R2R3 MYB transcription factor, GhMYB52 Like, in the development of the secondary cell wall in cotton fiber cells. Utilizing gene-editing technology to generate a loss-of-function mutant to clarify the role of GhMYB52 Like, we revealed that GhMYB52 Like does not directly contribute to cellulose synthesis in cotton fibers but instead represses a subset of lignin biosynthesis genes, establishing it as a lignin biosynthesis inhibitor. Concurrently, a substantial decrease in the lint index, a critical measure of cotton yield, was noted in parallel with an elevation in lignin levels. This study not only deepens our understanding of the molecular mechanisms underlying cotton fiber development but also offers new perspectives for the molecular improvement of other economically important crops and the enhancement of biomass energy utilization.


Assuntos
Fibra de Algodão , Regulação da Expressão Gênica de Plantas , Gossypium , Lignina , Proteínas de Plantas , Lignina/biossíntese , Gossypium/genética , Gossypium/metabolismo , Gossypium/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Parede Celular/metabolismo , Parede Celular/genética , Celulose/biossíntese , Celulose/metabolismo , Vias Biossintéticas
3.
Appl Microbiol Biotechnol ; 108(1): 324, 2024 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-38713211

RESUMO

Laccase, a copper-containing polyphenol oxidase, is an important green biocatalyst. In this study, Laccase Lcc5 was homologous recombinantly expressed in Coprinopsis cinerea and a novel strategy of silencing chitinase gene expression was used to enhance recombinant Lcc5 extracellular yield. Two critical chitinase genes, ChiEn1 and ChiE2, were selected by analyzing the transcriptome data of C. cinerea FA2222, and their silent expression was performed by RNA interference (RNAi). It was found that silencing either ChiEn1 or ChiE2 reduced sporulation and growth rate, and increased cell wall sensitivity, but had no significant effect on mycelial branching. Among them, the extracellular laccase activity of the ChiE2-silenced engineered strain Cclcc5-antiChiE2-5 and the control Cclcc5-13 reached the highest values (38.2 and 25.5 U/mL, respectively) at 250 and 150 rpm agitation speeds, corresponding to productivity of 0.35 and 0.19 U/mL·h, respectively, in a 3-L fermenter culture. Moreover, since Cclcc5-antiChiE2-5 could withstand greater shear forces, its extracellular laccase activity was 2.6-fold higher than that of Cclcc5-13 when the agitation speed was all at 250 rpm. To our knowledge, this is the first report of enhanced recombinant laccase production in C. cinerea by silencing the chitinase gene. This study will pave the way for laccase industrial production and accelerate the development of a C. cinerea high-expression system. KEY POINTS: • ChiEn1 and ChiE2 are critical chitinase genes in C. cinerea FA2222 genome. • Chitinase gene silencing enhanced the tolerance of C. cinerea to shear forces. • High homologous production of Lcc5 is achieved by fermentation in a 3-L fermenter.


Assuntos
Quitinases , Inativação Gênica , Lacase , Quitinases/genética , Quitinases/metabolismo , Quitinases/biossíntese , Lacase/genética , Lacase/metabolismo , Lacase/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Agaricales/genética , Agaricales/enzimologia , Fermentação , Interferência de RNA , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Micélio/genética , Micélio/crescimento & desenvolvimento , Micélio/enzimologia , Parede Celular/metabolismo , Parede Celular/genética
4.
J Mol Biol ; 436(10): 168570, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38604529

RESUMO

Cellular mRNA levels, particularly under stress conditions, can be finely regulated by the coordinated action of transcription and degradation processes. Elements of the 5'-3' mRNA degradation pathway, functionally associated with the exonuclease Xrn1, can bind to nuclear chromatin and modulate gene transcription. Within this group are the so-called decapping activators, including Pat1, Dhh1, and Lsm1. In this work, we have investigated the role of Pat1 in the yeast adaptive transcriptional response to cell wall stress. Thus, we demonstrated that in the absence of Pat1, the transcriptional induction of genes regulated by the Cell Wall Integrity MAPK pathway was significantly affected, with no effect on the stability of these transcripts. Furthermore, under cell wall stress conditions, Pat1 is recruited to Cell Wall Integrity-responsive genes in parallel with the RNA Pol II complex, participating both in pre-initiation complex assembly and transcriptional elongation. Indeed, strains lacking Pat1 showed lower recruitment of the transcription factor Rlm1, less histone H3 displacement at Cell Wall Integrity gene promoters, and impaired recruitment and progression of RNA Pol II. Moreover, Pat1 and the MAPK Slt2 occupied the coding regions interdependently. Our results support the idea that Pat1 and presumably other decay factors behave as transcriptional regulators of Cell Wall Integrity-responsive genes under cell wall stress conditions.


Assuntos
Parede Celular , Regulação Fúngica da Expressão Gênica , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Parede Celular/metabolismo , Parede Celular/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Sistema de Sinalização das MAP Quinases , Transcrição Gênica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Polimerase II/metabolismo , RNA Polimerase II/genética , Estabilidade de RNA , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Endorribonucleases/metabolismo , Endorribonucleases/genética , Proteínas de Ligação a RNA , Proteínas de Domínio MADS
5.
Dev Cell ; 59(10): 1345-1359.e6, 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38579721

RESUMO

The plant cell wall is a dynamic structure that plays an essential role in development, but the mechanism regulating cell wall formation remains poorly understood. We demonstrate that two transcription factors, SlERF.H5 and SlERF.H7, control cell wall formation and tomato fruit firmness in an additive manner. Knockout of SlERF.H5, SlERF.H7, or both genes decreased cell wall thickness, firmness, and cellulose contents in fruits during early development, especially in double-knockout lines. Overexpressing either gene resulted in thicker cell walls and greater fruit firmness with elevated cellulose levels in fruits but severely dwarf plants with lower gibberellin contents. We further identified that SlERF.H5 and SlERF.H7 activate the cellulose biosynthesis gene SlCESA3 but repress the gibberellin biosynthesis gene GA20ox1. Moreover, we identified a conserved LPL motif in these ERFs responsible for their activities as transcriptional activators and repressors, providing insight into how bifunctional transcription factors modulate distinct developmental processes.


Assuntos
Parede Celular , Frutas , Regulação da Expressão Gênica de Plantas , Giberelinas , Proteínas de Plantas , Solanum lycopersicum , Fatores de Transcrição , Solanum lycopersicum/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/crescimento & desenvolvimento , Giberelinas/metabolismo , Parede Celular/metabolismo , Parede Celular/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Frutas/metabolismo , Frutas/genética , Frutas/crescimento & desenvolvimento , Celulose/metabolismo , Celulose/biossíntese , Plantas Geneticamente Modificadas/metabolismo , Sequência Conservada , Motivos de Aminoácidos
6.
Int J Biol Macromol ; 268(Pt 1): 131559, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38631576

RESUMO

Expansins are important plant cell wall proteins. They can loosen and soften the cell walls and lead to wall extension and cell expansion. To investigate their role in wood formation and fiber elongation, the PagEXPA1 that highly expressed in cell differentiation and expansion tissues was cloned from 84K poplar (Populus alba × P. glandulosa). The subcellular localization showed that PagEXPA1 located in the cell wall and it was highly expressed in primary stems and young leaves. Compared with non-transgenic 84K poplar, overexpression of PagEXPA1 can promote plant-growth, lignification, and fiber cell elongation, while PagEXPA1 Cas9-editing mutant lines exhibited the opposite phenotype. Transcriptome analysis revealed that DEGs were mainly enriched in some important processes, which are associated with cell wall formation and cellulose synthesis. The protein interaction prediction and expression analysis showed that PagCDKB2:1 and PagEXPA1 might have an interaction relationship. The luciferase complementary assay and bimolecular fluorescence complementary assay validated that PagEXPA1 can combined with PagCDKB2;1. So they promoted the expansion of xylem vascular tissues and the development of poplar though participating in the regulation of cell division and differentiation by programming the cell-cycle. It provides good foundation for molecular breeding of fast-growing and high-quality poplar varieties.


Assuntos
Parede Celular , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Populus , Populus/genética , Populus/crescimento & desenvolvimento , Populus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Parede Celular/metabolismo , Parede Celular/genética , Plantas Geneticamente Modificadas , Perfilação da Expressão Gênica , Xilema/metabolismo , Xilema/genética , Desenvolvimento Vegetal/genética , Madeira/genética , Madeira/crescimento & desenvolvimento
7.
PLoS Genet ; 20(4): e1011234, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38598601

RESUMO

Peptidoglycan (PG) is the main component of the bacterial cell wall; it maintains cell shape while protecting the cell from internal osmotic pressure and external environmental challenges. PG synthesis is essential for bacterial growth and survival, and a series of PG modifications are required to allow expansion of the sacculus. Endopeptidases (EPs), for example, cleave the crosslinks between adjacent PG strands to allow the incorporation of newly synthesized PG. EPs are collectively essential for bacterial growth and must likely be carefully regulated to prevent sacculus degradation and cell death. However, EP regulation mechanisms are poorly understood. Here, we used TnSeq to uncover novel EP regulators in Vibrio cholerae. This screen revealed that the carboxypeptidase DacA1 (PBP5) alleviates EP toxicity. dacA1 is essential for viability on LB medium, and this essentiality was suppressed by EP overexpression, revealing that EP toxicity both mitigates, and is mitigated by, a defect in dacA1. A subsequent suppressor screen to restore viability of ΔdacA1 in LB medium identified hypomorphic mutants in the PG synthesis pathway, as well as mutations that promote EP activation. Our data thus reveal a more complex role of DacA1 in maintaining PG homeostasis than previously assumed.


Assuntos
Carboxipeptidases , Parede Celular , Endopeptidases , Peptidoglicano , Vibrio cholerae , Peptidoglicano/metabolismo , Vibrio cholerae/genética , Vibrio cholerae/metabolismo , Endopeptidases/genética , Endopeptidases/metabolismo , Carboxipeptidases/genética , Carboxipeptidases/metabolismo , Parede Celular/metabolismo , Parede Celular/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Epistasia Genética , Mutação
8.
Sheng Wu Gong Cheng Xue Bao ; 40(4): 1002-1016, 2024 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-38658144

RESUMO

Hemicellulose, as a primary component of plant cell walls, constitutes approximately one third of cell wall dry matter and ranks as the second abundant renewable biomass resource in the nature after cellulose. Hemicellulose is tightly cross-linked with cellulose, lignin and other components in the plant cell wall, leading to lignocellulose recalcitrance. However, precise genetic modifications of plant cell walls can significantly improve the saccharification efficiency of lignocellulose while ensuring normal plant growth and development. We comprehensively review the research progress in the structural distribution of hemicellulose in plant cell walls, the cross-linking between hemicellulose and other components of the cell wall, and the impact of hemicellulose modification on the saccharification efficiency of the cell wall, proving a reference for the genetic improvement of energy crops.


Assuntos
Parede Celular , Celulose , Lignina , Polissacarídeos , Parede Celular/metabolismo , Parede Celular/genética , Polissacarídeos/metabolismo , Lignina/metabolismo , Celulose/metabolismo , Plantas/genética , Plantas/metabolismo , Produtos Agrícolas/genética , Plantas Geneticamente Modificadas/genética
9.
Plant Sci ; 343: 112058, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38447913

RESUMO

The NF-Y gene family in plants plays a crucial role in numerous biological processes, encompassing hormone response, stress response, as well as growth and development. In this study, we first used bioinformatics techniques to identify members of the NF-YA family that may function in wood formation. We then used molecular biology techniques to investigate the role and molecular mechanism of PtrNF-YA6 in secondary cell wall (SCW) formation in Populus trichocarpa. We found that PtrNF-YA6 protein was localized in the nucleus and had no transcriptional activating activity. Overexpression of PtrNF-YA6 had an inhibitory effect on plant growth and development and significantly suppressed hemicellulose synthesis and SCW thickening in transgenic plants. Yeast one-hybrid and ChIP-PCR assays revealed that PtrNF-YA6 directly regulated the expression of hemicellulose synthesis genes (PtrGT47A-1, PtrGT8C, PtrGT8F, PtrGT43B, PtrGT47C, PtrGT8A and PtrGT8B). In conclusion, PtrNF-YA6 can inhibit plant hemicellulose synthesis and SCW thickening by regulating the expression of downstream SCW formation-related target genes.


Assuntos
Populus , Populus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Madeira/genética , Parede Celular/genética , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
10.
Ann Bot ; 133(7): 953-968, 2024 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-38366549

RESUMO

BACKGROUND AND AIMS: Secondary cell wall (SCW) thickening is a major cellular developmental stage determining wood structure and properties. Although the molecular regulation of cell wall deposition during tracheary element differentiation has been well established in primary growth systems, less is known about the gene regulatory processes involved in the multi-layered SCW thickening of mature trees. METHODS: Using third-generation [long-read single-molecule real-time (SMRT)] and second-generation [short-read sequencing by synthesis (SBS)] sequencing methods, we established a Pinus bungeana transcriptome resource with comprehensive functional and structural annotation for the first time. Using these approaches, we generated high spatial resolution datasets for the vascular cambium, xylem expansion regions, early SCW thickening, late SCW thickening and mature xylem tissues of 71-year-old Pinus bungeana trees. KEY RESULTS: A total of 79 390 non-redundant transcripts, 31 808 long non-coding RNAs and 5147 transcription factors were annotated and quantified in different xylem tissues at all growth and differentiation stages. Furthermore, using this high spatial resolution dataset, we established a comprehensive transcriptomic profile and found that members of the NAC, WRKY, SUS, CESA and LAC gene families are major players in early SCW formation in tracheids, whereas members of the MYB and LBD transcription factor families are highly expressed during late SCW thickening. CONCLUSIONS: Our results provide new molecular insights into the regulation of multi-layered SCW thickening in conifers. The high spatial resolution datasets provided can serve as important gene resources for improving softwoods.


Assuntos
Parede Celular , Pinus , Xilema , Parede Celular/genética , Parede Celular/metabolismo , Pinus/genética , Pinus/crescimento & desenvolvimento , Xilema/genética , Xilema/crescimento & desenvolvimento , Transcriptoma , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Madeira/genética , Madeira/crescimento & desenvolvimento , Madeira/anatomia & histologia
11.
Fungal Genet Biol ; 171: 103876, 2024 03.
Artigo em Inglês | MEDLINE | ID: mdl-38367799

RESUMO

Colletotrichum graminicola, the causal agent of maize leaf anthracnose and stalk rot, differentiates a pressurized infection cell called an appressorium in order to invade the epidermal cell, and subsequently forms biotrophic and necrotrophic hyphae to colonize the host tissue. While the role of force in appressorial penetration is established (Bechinger et al., 1999), the involvement of cell wall-degrading enzymes (CWDEs) in this process and in tissue colonization is poorly understood, due to the enormous number and functional redundancy of these enzymes. The serine/threonine protein kinase gene SNF1 identified in Sucrose Non-Fermenting yeast mutants mediates de-repression of catabolite-repressed genes, including many genes encoding CWDEs. In this study, we identified and functionally characterized the SNF1 homolog of C. graminicola. Δsnf1 mutants showed reduced vegetative growth and asexual sporulation rates on media containing polymeric carbon sources. Microscopy revealed reduced efficacies in appressorial penetration of cuticle and epidermal cell wall, and formation of unusual medusa-like biotrophic hyphae by Δsnf1 mutants. Severe and moderate virulence reductions were observed on intact and wounded leaves, respectively. Employing RNA-sequencing we show for the first time that more than 2,500 genes are directly or indirectly controlled by Snf1 in necrotrophic hyphae of a plant pathogenic fungus, many of which encode xylan- and cellulose-degrading enzymes. The data presented show that Snf1 is a global regulator of gene expression and is required for full virulence.


Assuntos
Colletotrichum , Zea mays , Zea mays/genética , Virulência/genética , Parede Celular/genética , Parede Celular/metabolismo , Saccharomyces cerevisiae/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Doenças das Plantas/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo
12.
Plant Physiol ; 194(4): 2472-2490, 2024 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-38217865

RESUMO

LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKEs (LBDs/ASLs) are plant-specific transcription factors that function downstream of auxin-regulated lateral root (LR) formation. Our previous research found that PpLBD16 positively regulates peach (Prunus persica) LR formation. However, the downstream regulatory network and target genes of PpLBD16 are still largely unknown. Here, we constructed a PpLBD16 homologous overexpression line and a PpLBD16 silenced line. We found that overexpressing PpLBD16 promoted peach root initiation, while silencing PpLBD16 inhibited peach root formation. Through RNA sequencing (RNA-seq) analysis of roots from PpLBD16 overexpression and silenced lines, we discovered that genes positively regulated by PpLBD16 were closely related to cell wall synthesis and degradation, ion/substance transport, and ion binding and homeostasis. To further detect the binding motifs and potential target genes of PpLBD16, we performed DNA-affinity purification sequencing (DAP-seq) analysis in vitro. PpLBD16 preferentially bound to CCNGAAANNNNGG (MEME-1), [C/T]TTCT[C/T][T/C] (MEME-2), and GCGGCGG (ABR1) motifs. By combined analysis of RNA-seq and DAP-seq data, we screened candidate target genes for PpLBD16. We demonstrated that PpLBD16 bound and activated the cell wall modification-related genes EXPANSIN-B2 (PpEXPB2) and SUBTILISIN-LIKE PROTEASE 1.7 (PpSBT1.7), the ion transport-related gene CYCLIC NUCLEOTIDE-GATED ION CHANNEL 1 (PpCNGC1) and the polyphenol oxidase (PPO)-encoding gene PpPPO, thereby controlling peach root organogenesis and promoting LR formation. Moreover, our results displayed that PpLBD16 and its target genes are involved in peach LR primordia development. Overall, this work reveals the downstream regulatory network and target genes of PpLBD16, providing insights into the molecular network of LBD16-mediated LR development.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Prunus persica , Fatores de Transcrição/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Prunus persica/genética , Prunus persica/metabolismo , Regulação da Expressão Gênica de Plantas , Transporte de Íons , Parede Celular/genética , Parede Celular/metabolismo , Raízes de Plantas/metabolismo , Ácidos Indolacéticos/metabolismo
13.
Arch Microbiol ; 206(1): 53, 2024 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-38180563

RESUMO

The A. sendaiensis PA2 is a polyextremophile bacterium. In this study, we analyze the A. sendaiensis PA2 genome. The genome was assembled and annotated. The A. sendaiensis PA2 genome structure consists of a 2,956,928 bp long chromosome and 62.77% of G + C content. 3056 CDSs were predicted, and 2921 genes were assigned to a putative function. The ANIm and ANIb value resulted in 97.17% and 96.65%, the DDH value was 75.5%, and the value of TETRA (Z-score) was 0.98. Comparative genomic analyses indicated that three systems are enriched in A. sendaiensis PA2. This strain has phenotypic changes in cell wall during batch culture at 65 °C, pH 5.0 and without carbon and nitrogen source. The presence of unique genes of cell wall and sporulation subsystem could be related to the adaptation of A. sendaiensis PA2 to hostile conditions.


Assuntos
Alicyclobacillus , Temperatura , Parede Celular/genética , Concentração de Íons de Hidrogênio
14.
Int J Mol Sci ; 25(2)2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38256004

RESUMO

Crown gall disease (Agrobacterium tumefaciens), crown/root rot disease (Phytophthora spp.), root lesion disease (Pratylenchus vulnus) and tree vigor are key traits affecting the productivity and quality of walnuts in California. Unchallenged hybrid rootstocks were analyzed by RNA-seq to examine pre-formed factors affecting these traits. Enrichment analysis of the differentially expressed genes revealed that the increased expression of cell wall biogenesis-related genes plays a key role in susceptibility to A. tumefaciens, susceptibility to Phytophthora spp. and increased vigor. Analysis of the predicted subcellular loci of the encoded proteins revealed that many gene products associated with vigor and susceptibility were targeted to the plasma membrane and extracellular space, connecting these traits to sustaining barrier function. We observed that RNA processing and splicing, along with predicted nuclear targeting, were associated with resistance to A. tumefaciens, resistance to Phytophthora spp. and low vigor. Four genes within the J. microcarpa QTL region for resistance to A. tumefaciens and Phytophthora spp. were represented among our transcripts, with two of the genes being differentially expressed in association with resistance to A. tumefaciens and decreased vigor. No differential expression related to Phytophthora spp. or P. vulnus resistance was observed in this region. Additionally, the J. microcarpa haplotype expressed more transcripts associated with resistance to A. tumefaciens, Phytophthora spp. and low vigor, but not P. vulnus, than the J. regia haplotype. We also report unique and shared hormone and defense responses associated with each trait. This research suggests a link between cell wall biogenesis, vigor and critical root diseases of walnut.


Assuntos
Juglans , Phytophthora , Juglans/genética , Perfilação da Expressão Gênica , Transcriptoma , Nozes , Parede Celular/genética
15.
Int J Biol Macromol ; 260(Pt 2): 129583, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38242409

RESUMO

Cell wall synthesis and cell division are two closely linked pathways in a bacterial cell which distinctly influence the growth and survival of a bacterium. This requires an appreciable coordination between the two processes, more so, in case of mycobacteria with an intricate multi-layered cell wall structure. In this study, we investigated a conserved gene cluster using CRISPR-Cas12 based gene silencing technology to show that knockdown of most of the genes in this cluster leads to growth defects. Investigating conserved genes is important as they likely perform vital cellular functions and the functional insights on such genes can be extended to other mycobacterial species. We characterised one of the genes in the locus, MSMEG_0311. The repression of this gene not only imparts severe growth defect but also changes colony morphology. We demonstrate that the protein preferentially localises to the polar region and investigate its influence on the polar growth of the bacillus. A combination of permeability and drug susceptibility assay strongly suggests a cell wall associated function of this gene which is also corroborated by transcriptomic analysis of the knockdown where a number of cell wall associated genes, particularly iniA and sigF regulon get altered. Considering the gene is highly conserved across mycobacterial species and appears to be essential for growth, it may serve as a potential drug target.


Assuntos
Mycobacterium tuberculosis , Mycobacterium , Mycobacterium smegmatis/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Mycobacterium/genética , Mycobacterium/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Divisão Celular , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/metabolismo
16.
Biol Futur ; 74(4): 519-527, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37964139

RESUMO

The chaperonin TRiC/CCT is cytosolic cylindrical complex of 16 subunits encoded by eight essential genes CCT1-8. It contributes to folding 10% of cellular polypeptides in yeast. The strain carrying substitution point mutation G412E in the equatorial domain of Cct7p resulted in the improper folding of substrates. In this study, the Cct7p mutant exhibited sensitivity to non-optimal growth temperatures and cell wall stressors. Heat shock is known to disrupt cell wall and protein stability in budding yeast. Mitogen-activated protein kinase-mediated cell wall integrity pathway gets activated to compensate the perturbed cell wall. Overexpression of the PKC1 and SLT2 genes of MAPK signaling pathway in mutant rescued the growth and cell division defects. Additionally, the genes of the CWI pathway such as SED1, GFA1, PIR1, and RIM21 are down-regulated. The Cct7p mutant strain (G412E) is unable to withstand the heat stress due to the underlying defects in protein folding and cell wall maintenance. Taken together, our results strongly indicate the interaction between CCT and cell wall integrity pathway.


Assuntos
Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Resposta ao Choque Térmico/genética
17.
J Agric Food Chem ; 71(49): 19568-19580, 2023 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-38019936

RESUMO

Septins are a conserved group of GTP-binding proteins found in all eukaryotes and are the fourth-most abundant cytoskeletal proteins. Septins of some pathogenic fungi are involved in morphological changes related to infection. Our previous studies have identified four core septins (StSep1-4) in Setosphaeria turcica, the causal agent of northern corn leaf blight, while only StSep4 is significantly upregulated during the invasive process. We therefore used forchlorfenuron (FCF), the specific inhibitor of septin, and ΔStSep4 knockout mutants to further clarify the role of septins in S. turcica pathogenicity. FCF treatment caused a dose-dependent reduction in S. turcica colony growth, delayed the formation of infection structures, and reduced the penetration ability. ΔStSep4 knockout mutants displayed abnormal mycelium morphology, slow mycelial growth, conidiation deficiency, delayed appressorium development, and weakened pathogenicity. StSep4 deletion also broke cell wall integrity, altered chitin distribution, decreased the melanin content, and disrupted normal nuclear localization. A transcriptomic comparison revealed that genes differentially expressed between ΔStSep4 and WT were enriched in terms of ribosomes, protein translation, membrane components, and transmembrane transport activities. Our results demonstrate that StSep4 is required for morphology and pathogenicity in S. turcica, making it a promising target for the development of novel fungicides.


Assuntos
Septinas , Fatores de Virulência , Septinas/genética , Septinas/metabolismo , Virulência , Parede Celular/genética , Parede Celular/metabolismo
18.
PLoS One ; 18(11): e0293228, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38011112

RESUMO

Translation initiation is a complex and highly regulated process that represents an important mechanism, controlling gene expression. eIF2A was proposed as an alternative initiation factor, however, its role and biological targets remain to be discovered. To further gain insight into the function of eIF2A in Saccharomyces cerevisiae, we identified mRNAs associated with the eIF2A complex and showed that 24% of the most enriched mRNAs encode proteins related to cell wall biogenesis and maintenance. In agreement with this result, we showed that an eIF2A deletion sensitized cells to cell wall damage induced by calcofluor white. eIF2A overexpression led to a growth defect, correlated with decreased synthesis of several cell wall proteins. In contrast, no changes were observed in the transcriptome, suggesting that eIF2A controls the expression of cell wall-related proteins at a translational level. The biochemical characterization of the eIF2A complex revealed that it strongly interacts with the RNA binding protein, Ssd1, which is a negative translational regulator, controlling the expression of cell wall-related genes. Interestingly, eIF2A and Ssd1 bind several common mRNA targets and we found that the binding of eIF2A to some targets was mediated by Ssd1. Surprisingly, we further showed that eIF2A is physically and functionally associated with the exonuclease Xrn1 and other mRNA degradation factors, suggesting an additional level of regulation. Altogether, our results highlight new aspects of this complex and redundant fine-tuned regulation of proteins expression related to the cell wall, a structure required to maintain cell shape and rigidity, providing protection against harmful environmental stress.


Assuntos
Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , RNA Mensageiro/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Expressão Gênica , Regulação Fúngica da Expressão Gênica
19.
J Mol Evol ; 91(6): 761-772, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37979044

RESUMO

Much evidence exists suggesting the presence of genetic functional diversification in plants, though literature associated with the role of functional diversification in the evolution of the plant secondary cell wall (SCW) has sparsely been compiled and reviewed in a recent context. This review aims to elucidate, through the examination of gene phylogenies associated with its biosynthesis and maintenance, the role of functional diversification in shaping the critical, dynamic, and characteristic organelle, the secondary cell wall. It will be asserted that gene families resulting from gene duplication and subsequent functional divergence are present and are heavily involved in SCW biosynthesis and maintenance. Furthermore, diversification will be presented as a significant driver behind the evolution of the many functional characteristics of the SCW. The structure and function of the plant cell wall and its constituents will first be explored, followed by a discussion on the phenomenon of gene duplication and the resulting genetic functional divergence that can emerge. Finally, the major constituents of the SCW and their individual relationships with duplication and divergence will be reviewed to the extent of current knowledge on the subject.


Assuntos
Parede Celular , Plantas , Plantas/genética , Parede Celular/genética , Duplicação Gênica , Regulação da Expressão Gênica de Plantas
20.
Mol Biol Rep ; 50(12): 10509-10524, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37921982

RESUMO

The focus is now on harnessing energy from green sources through sustainable technology to minimize environmental pollution. Several crop residues including rice and wheat straw are having enormous potential to be used as lignocellulosic source material for bioenergy production. The lignocellulosic feedstock is primarily composed of cellulose, hemicellulose, and lignin cell wall polymers. The hemicellulose and lignin polymers induce crosslinks in the cell wall, by firmly associating with cellulose microfibrils, and thereby, denying considerable access of cellulose to cellulase enzymes. This issue has been addressed by various researchers through downregulating several genes associated in monolignol biosynthesis in Arabidopsis, Poplar, Rice and Switchgrass to increase ethanol recovery. Similarly, xylan biosynthetic genes are also targeted to genetically culminate its accumulation in the secondary cell walls. Regulation of cellulose synthases (CesA) proves to be an effective tool in addressing the negative impact of these two factors. Modification in the expression of cellulose synthase aids in reducing cellulose crystallinity as well as polymerisation degree which in turn increases ethanol recovery. The engineered bioenergy crops and various fungal strains with state of art biomass processing techniques presents the most recent integrative biotechnology model for cost effective green fuels generation along with production of key value-added products with minuscule disturbances in the environment. Plant breeding strategies utilizing the existing variability for biomass traits will be key in developing dual purpose varieties. For this purpose, reorientation of conventional breeding techniques for incorporating useful biomass traits will be effective.


Assuntos
Arabidopsis , Oryza , Lignina/metabolismo , Melhoramento Vegetal , Celulose/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Polímeros , Oryza/genética , Oryza/metabolismo , Arabidopsis/metabolismo , Etanol/metabolismo , Biomassa
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