RESUMEN
BACKGROUND: Recent large-scale whole genome sequencing efforts in birds have elucidated broad patterns of avian phylogeny and genome evolution. However, despite the great interest in economically important phasianids like Gallus gallus (Red Junglefowl, the progenitor of the chicken), we know little about the genomes of closely related species. Gallus gallus is highly sexually dichromatic and polygynous, but its sister genus, Bambusicola, is smaller, sexually monomorphic, and monogamous with biparental care. We sequenced the genome of Bambusicola thoracicus (Chinese Bamboo Partridge) using a single insert library to test hypotheses about genome evolution in galliforms. Selection acting at the phenotypic level could result in more evidence of positive selection in the Gallus genome than in Bambusicola. However, the historical range size of Bambusicola was likely smaller than Gallus, and demographic effects could lead to higher rates of nonsynonymous substitution in Bambusicola than in Gallus. RESULTS: We generated a genome assembly suitable for evolutionary analyses. We examined the impact of selection on coding regions by examining shifts in the average nonsynonymous to synonymous rate ratio (dN/dS) and the proportion of sites subject to episodic positive selection. We observed elevated dN/dS in Bambusicola relative to Gallus, which is consistent with our hypothesis that demographic effects may be important drivers of genome evolution in Bambusicola. We also demonstrated that alignment error can greatly inflate estimates of the number of genes that experienced episodic positive selection and heterogeneity in dN/dS. However, overall patterns of molecular evolution were robust to alignment uncertainty. Bambusicola thoracicus has higher estimates of heterozygosity than Gallus gallus, possibly due to migration events over the past 100,000 years. CONCLUSIONS: Our results emphasized the importance of demographic processes in generating the patterns of variation between Bambusicola and Gallus. We also demonstrated that genome assemblies generated using a single library can provide valuable insights into avian evolutionary history and found that it is important to account for alignment uncertainty in evolutionary inferences from draft genomes.
Asunto(s)
Evolución Molecular , Galliformes/genética , Genómica , Animales , Ontología de Genes , Heterocigoto , Anotación de Secuencia MolecularRESUMEN
Genome projects have revolutionized our understanding of both molecular biology and evolution, but there has been a limited collection of genomic data from reptiles. This is surprising given the pivotal position of reptiles in vertebrate phylogeny and the potential utility of information from reptiles for understanding a number of biological phenomena, such as sex determination. Although there are many potential uses for genomic data, one important and useful approach is phylogenomics. Here we report cDNA sequences for the c-Jun(JUN) and DJ-1(PARK7) proto-oncogenes from 3 reptiles (the American alligator, Nile crocodile, and Florida red-belly turtle), show that both genes are expressed in the alligator, and integrate them into analyses of their homologs from other organisms. With these taxa it was possible to conduct analyses that include all major vertebrate lineages. Analyses of c-Jun revealed an unexpected but well-supported frog-turtle clade while analyses of DJ-1 revealed a topology largely congruent with expectation based upon other data. The conflict between the c-Jun topology and expectation appears to reflect the overlap between c-Jun and a CpG island in most taxa, including crocodilians. This CpG island is absent in the frog and turtle, and convergence in base composition appears to be at least partially responsible for the signal uniting these taxa. Noise reduction approaches can eliminate the unexpected frog-turtle clade, demonstrating that multiple signals are present in the c-Jun alignment. We used phylogenetic methods to visualize these signals; we suggest that examining both historical and non-historical signals will prove important for phylogenomic analyses.
Asunto(s)
Caimanes y Cocodrilos/genética , Genes jun/genética , Proteínas Oncogénicas/genética , Filogenia , Tortugas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Secuencia Conservada , Islas de CpG/genética , Genómica , Péptidos y Proteínas de Señalización Intracelular/genética , Datos de Secuencia Molecular , Proteína Desglicasa DJ-1 , ARN Mensajero/genética , Transducción de Señal/genéticaRESUMEN
Mutations in the transparent testa (tt) loci abolish pigment production in Arabidopsis seed coats. The TT4, TT5, and TT3 loci encode chalcone synthase, chalcone isomerase, and dihydroflavonol 4-reductase, respectively, which are essential for anthocyanin accumulation and may form a macromolecular complex. Here, we show that the products of the maize (Zea mays) C2, CHI1, and A1 genes complement Arabidopsis tt4, tt5, and tt3 mutants, restoring the ability of these mutants to accumulate pigments in seed coats and seedlings. Overexpression of the maize genes in wild-type Arabidopsis seedlings does not result in increased anthocyanin accumulation, suggesting that the steps catalyzed by these enzymes are not rate limiting in the conditions assayed. The expression of the maize A1 gene in the flavonoid 3' hydroxylase Arabidopsis tt7 mutant resulted in an increased accumulation of pelargonidin. We conclude that enzymes involved in secondary metabolism can be functionally exchangeable between plants separated by large evolutionary distances. This is in sharp contrast to the notion that the more relaxed selective constrains to which secondary metabolic pathways are subjected is responsible for the rapid divergence of the corresponding enzymes.
Asunto(s)
Aciltransferasas/genética , Oxidorreductasas de Alcohol/genética , Arabidopsis/enzimología , Flavonoides/biosíntesis , Liasas Intramoleculares/genética , Zea mays/enzimología , Aciltransferasas/metabolismo , Oxidorreductasas de Alcohol/metabolismo , Secuencia de Aminoácidos , Antocianinas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Flavonoides/química , Flavonoides/genética , Flavonoides/metabolismo , Regulación de la Expresión Génica de las Plantas , Prueba de Complementación Genética , Liasas Intramoleculares/metabolismo , Datos de Secuencia Molecular , Mutación , Proteínas de Plantas/metabolismo , Estructuras de las Plantas/metabolismo , Plantas Modificadas Genéticamente , Semillas/metabolismo , Zea mays/genética , Zea mays/metabolismoAsunto(s)
Evolución Molecular , Filogenia , Modelos Genéticos , Tamaño de la Muestra , Procesos EstocásticosRESUMEN
Proteins are often characterized by the presence of multiple domains, which make specific contributions to their cellular function. While the gain of domains in proteins by duplication and shuffling is well established, domain loss is poorly documented. Here, we provide evidence that domain loss has played an important role in the evolution of protein architecture and function by demonstrating that fungal Zuotin proteins evolved from MIDA1-like proteins, present in animals and plants, by complete loss of the carboxyl-terminal MYB domains. Phylogenetic analyses of the DnaJ motif (the J domain) present in both Zuotin and MIDA1 proteins were complicated by the limited length and profound differences in evolutionary rates exhibited by this domain. To rigorously examine J domain phylogeny, we combined the nonparametric bootstrap with Monte Carlo simulation. This method, which we have designated the resampled parametric bootstrap, allowed us to assess type I and type II error associated with these analyses. These results revealed significant support for domain loss rather than domain gain or gene loss involving paralogs. The absence of sequences related to the MIDA1 MYB domains in Saccharomyces cerevisiae further indicates that the domains have been completely lost, consistent with known functional differences between Zuotin and MIDA1 proteins. These analyses suggest that the description of additional examples of complete domain loss may provide a method to identify orthologous proteins exhibiting functional differences using genomic sequence data.
Asunto(s)
Proteínas de Unión al ADN/genética , Evolución Molecular , Proteínas Fúngicas/genética , Proteínas de Saccharomyces cerevisiae , Secuencia de Aminoácidos , Animales , Arabidopsis/genética , Proteínas de Unión al ADN/química , Proteínas Fúngicas/química , Ratones , Modelos Genéticos , Chaperonas Moleculares , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Estructura Terciaria de Proteína , Saccharomyces cerevisiae/genética , Eliminación de Secuencia , Homología de Secuencia de AminoácidoRESUMEN
The entire mitochondrial DNA control region (mtDNA CR) and cytochrome b (cyt b) genes were sequenced in 10 of the 11 extant species of gallopheasants (Lophura). The cyt b from L. diardi and L. ignita showed unusual leucine-coding codons at the expected terminal 3' end of the gene. Presence of conserved functional motifs in the inferred amino acid sequences, conserved secondary structures of the flanking tRNA(Pro) and tRNA(Thr), and Southern hybridization concordantly suggest that these cyt b represent functional mitochondrial genes and not nuclear transpositions. Functional stop codons can be generated by RNA editing of the primary transcripts from these sequences. Despite strong site and domain substitution rate heterogeneity, CR and cyt b diverged at similar rates, on average, and expressed congruent phylogenetic signals. Phylogenetic analyses of the concatenated sequences split Lophura into five clades including (1) L. bulweri, (2) L. diardi-L. ignita, (3) L. erythrophthalma-L. inornata, (4) L. leucomelanos-L. nycthemera, and (5) L. swinhoii-L. edwardsi-L. hatinhensis. Basal relationships among these clades, which include species distributed in continental South East Asia and the Sundaland archipelago, were weakly resolved, suggesting the occurrence of rapid cladogenic events in the early evolutionary history of Lophura. A conventional calibration of mtDNA sequence divergence indicates a mid to late Pliocene evolution of the main clades in Lophura, which could have diversified in allopatry in continental South East Asia. Sundaland could have been colonized lately and independently by the different clades. Consequently, cyclic changes in late Pleistocene climate and landscape might not have increased rates of speciation in genus Lophura in Sundaland.
Asunto(s)
Aves/genética , Grupo Citocromo b/genética , ADN Mitocondrial/genética , Evolución Molecular , Filogenia , Animales , Secuencia de Bases , Aves/clasificación , ADN Mitocondrial/química , Variación Genética , Datos de Secuencia Molecular , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
We report the analysis of a 36-kbp region of the Neurospora crassa genome, which contains homologs of two closely linked stationary phase genes, SNZ1 and SNO1, from Saccharomyces cerevisiae. Homologs of SNZ1 encode extremely highly conserved proteins that have been implicated in pyridoxine (vitamin B6) metabolism in the filamentous fungi Cercospora nicotianae and in Aspergillus nidulans. In N. crassa, SNZ and SNO homologs map to the region occupied by pdx-1 (pyridoxine requiring), a gene that has been known for several decades, but which was not sequenced previously. In this study, pyridoxine-requiring mutants of N. crassa were found to possess mutations that disrupt conserved regions in either the SNZ or SNO homolog. Previously, nearly all of these mutants were classified as pdx-1. However, one mutant with a disrupted SNO homolog was at one time designated pdx-2. It now appears appropriate to reserve the pdx-1 designation for the N. crassa SNZ homolog and pdx-2 for the SNO homolog. We further report annotation of the entire 36,030-bp region, which contains at least 12 protein coding genes, supporting a previous conclusion of high gene densities (12,000-13,000 total genes) for N. crassa. Among genes in this region other than SNZ and SNO homologs, there was no evidence of shared function. Four of the genes in this region appear to have been lost from the S. cerevisiae lineage.
Asunto(s)
Proteínas Fúngicas/genética , Genoma Fúngico , Neurospora crassa/genética , Piridoxina/metabolismo , Proteínas de Saccharomyces cerevisiae , Clonación Molecular , Cósmidos , Biblioteca de Genes , Ligamiento Genético , Modelos Genéticos , Mutación , Sistemas de Lectura Abierta , Fenotipo , Análisis de Secuencia de ADNRESUMEN
We report a large-scale comparison of sequence data from the filamentous fungus Neurospora crassa with the complete genome sequence of Saccharomyces cerevisiae. N. crassa is considerably more morphologically and developmentally complex than S. cerevisiae. We found that N. crassa has a much higher proportion of "orphan" genes than S. cerevisiae, suggesting that its morphological complexity reflects the acquisition or maintenance of novel genes, consistent with its larger genome. Our results also indicate the loss of specific genes from S. cerevisiae. Surprisingly, some of the genes lost from S. cerevisiae are involved in basic cellular processes, including translation and ion (especially calcium) homeostasis. Horizontal gene transfer from prokaryotes appears to have played a relatively modest role in the evolution of the N. crassa genome. Differences in the overall rate of molecular evolution between N. crassa and S. cerevisiae were not detected. Our results indicate that the current public sequence databases have fairly complete samples of gene families with ancient conserved regions, suggesting that further sequencing will not substantially change the proportion of genes with homologs among distantly related groups. Models of the evolution of fungal genomes compatible with these results, and their functional implications, are discussed.
Asunto(s)
ADN de Hongos/análisis , Eliminación de Gen , Genes Fúngicos/genética , Neurospora crassa/genética , Saccharomyces cerevisiae/genética , Aspergillus nidulans/genética , Bases de Datos Factuales , Evolución Molecular , Etiquetas de Secuencia Expresada , Proteínas Fúngicas/biosíntesis , Proteínas Fúngicas/genética , Proteínas Fúngicas/aislamiento & purificación , Variación Genética/genética , Genoma Fúngico , Homología de Secuencia de Ácido NucleicoAsunto(s)
Evolución Molecular , Genes de Plantas/genética , Familia de Multigenes/genética , Plantas/genética , Proteínas Proto-Oncogénicas c-myb/genética , Secuencia de Aminoácidos , Animales , Datos de Secuencia Molecular , Filogenia , Proteínas Proto-Oncogénicas c-myb/química , Homología de Secuencia de AminoácidoRESUMEN
Transcription factors containing the Myb-homologous DNA-binding domain are widely found in eukaryotes. In plants, R2R3 Myb-domain proteins are involved in the control of form and metabolism. The Arabidopsis genome harbors >100 R2R3 Myb genes, but few have been found in monocots, animals, and fungi. Using RT-PCR from different maize organs, we cloned 480 fragments corresponding to a 42-44 residue-long sequence spanning the region between the conserved DNA-recognition helices (Myb(BRH)) of R2R3 Myb domains. We determined that maize expresses >80 different R2R3 Myb genes, and evolutionary distances among maize Myb(BRH) sequences indicate that most of the amplification of the R2R3 Myb gene family occurred after the origin of land plants but prior to the separation of monocots and dicots. In addition, evidence is provided for the very recent duplication of particular classes of R2R3 Myb genes in the grasses. Together, these findings render a novel line of evidence for the amplification of the R2R3 Myb gene family in the early history of land plants and suggest that maize provides a possible model system to examine the hypothesis that the expansion of Myb genes is associated with the regulation of novel plant cellular functions.
Asunto(s)
Proteínas de Unión al ADN/genética , Evolución Molecular , Amplificación de Genes/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Proteínas Proto-Oncogénicas c-myb , Zea mays/genética , Secuencia de Aminoácidos , Animales , Proteínas de Arabidopsis , Clonación Molecular , Codón/genética , Proteínas de Unión al ADN/química , Duplicación de Gen , Genes de Plantas/fisiología , Variación Genética/genética , Humanos , Datos de Secuencia Molecular , Familia de Multigenes/genética , Mutación/genética , Filogenia , Proteínas de Plantas/química , Estructuras de las Plantas/genética , Alineación de SecuenciaRESUMEN
Opsins are a class of retinal-binding, seven transmembrane helix proteins that function as light-responsive ion pumps or sensory receptors. Previously, genes encoding opsins had been identified in animals and the Archaea but not in fungi or other eukaryotic microorganisms. Here, we report the identification and mutational analysis of an opsin gene, nop-1, from the eukaryotic filamentous fungus Neurospora crassa. The nop-1 amino acid sequence predicts a protein that shares up to 81.8% amino acid identity with archaeal opsins in the 22 retinal binding pocket residues, including the conserved lysine residue that forms a Schiff base linkage with retinal. Evolutionary analysis revealed relatedness not only between NOP-1 and archaeal opsins but also between NOP-1 and several fungal opsin-related proteins that lack the Schiff base lysine residue. The results provide evidence for a eukaryotic opsin family homologous to the archaeal opsins, providing a plausible link between archaeal and visual opsins. Extensive analysis of Deltanop-1 strains did not reveal obvious defects in light-regulated processes under normal laboratory conditions. However, results from Northern analysis support light and conidiation-based regulation of nop-1 gene expression, and NOP-1 protein heterologously expressed in Pichia pastoris is labeled by using all-trans [3H]retinal, suggesting that NOP-1 functions as a rhodopsin in N. crassa photobiology.
Asunto(s)
Archaea/genética , Proteínas Portadoras/química , Proteínas Portadoras/genética , Evolución Molecular , Neurospora crassa/genética , Filogenia , Retinaldehído/metabolismo , Rodopsina/genética , Secuencia de Aminoácidos , Archaea/clasificación , Bases de Datos Factuales , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Hongos/clasificación , Hongos/genética , Modelos Moleculares , Datos de Secuencia Molecular , Neurospora crassa/efectos de los fármacos , Neurospora crassa/crecimiento & desarrollo , Oligomicinas/farmacología , Estructura Secundaria de Proteína , Rodopsina/químicaRESUMEN
Cytochrome b and D-loop nucleotide sequences were used to study patterns of molecular evolution and phylogenetic relationships between the pheasants and the partridges, which are thought to form two closely related monophyletic galliform lineages. Our analyses used 34 complete cytochrome b and 22 partial D-loop sequences from the hypervariable domain I of the D-loop, representing 20 pheasant species (15 genera) and 12 partridge species (5 genera). We performed parsimony, maximum likelihood, and distance analyses to resolve these phylogenetic relationships. In this data set, transversion analyses gave results similar to those of global analyses. All of our molecular phylogenetic analyses indicated that the pheasants and partridges arose through a rapid radiation, making it difficult to establish higher level relationships. However, we were able to establish six major lineages containing pheasant and partridge taxa, including one lineage containing both pheasants and partridges (Gallus, Bambusicola and Francolinus). This result, supported by maximum likelihood tests, indicated that the pheasants and partridges do not form independent monophyletic lineages.
Asunto(s)
Aves/genética , Evolución Molecular , Filogenia , Animales , Secuencia de Bases , Grupo Citocromo b/genética , ADN Mitocondrial/química , ADN Mitocondrial/genética , Mitocondrias/genética , Datos de Secuencia Molecular , Alineación de Secuencia , Homología de Secuencia de Ácido NucleicoRESUMEN
Annexin homologues have been found in animals, plants, and distinct protist lineages. We report the identification of the first fungal annexin, encoded by the anx14 gene of the filamentous ascomycete Neurospora crassa. Annexins have a complex evolutionary history and exhibit a large number of gene duplications and gene losses in various taxa, including the complete loss of annexin sequences from another ascomycete, the budding yeast Saccharomyces cerevisiae. Surprisingly, the N. crassa annexin homologue is most closely related to the annexin homologue of the slime mold Dictyostelium discoideum, suggesting a phylogenetic link between cellular slime molds and true fungi. Both of these annexin homologues are closely related to the family of annexin homologues present in animals, an observation consistent with the existence of the animal-fungal clade. These data further suggest that the gene duplications that generated the family of annexin sequences present in animals, fungi, and slime molds began prior to the divergence of these taxa.
Asunto(s)
Anexinas/genética , Dictyostelium/genética , Evolución Molecular , Duplicación de Gen , Neurospora crassa/genética , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN Complementario , Humanos , Datos de Secuencia Molecular , FilogeniaRESUMEN
Afropavo congensis, the Congo peafowl, has long fascinated ornithologists because of its uncertain phylogenetic position and unusual geographic distribution. While some researchers have placed Afropavo as a sister taxon to the true peafowl, Pavo species, others have suggested relationships with the guineafowl or an Old World partridge, Francolinus. These divergent opinions are due, at least in part, to (i) the unique morphological characteristics, lack of elaborate ornamentation, and monogamous mating system in Afropavo which differentiates it from Pavo; and (ii) the restricted distribution of Afropavo in Zaire, which is far removed from the Asian distribution of all other pheasant species. We obtained complete cytochrome b and partial D-loop sequences of Afropavo and compared them to Pavo, guineafowl, Francolinus and other galliform taxa. Our results strongly support a close relationship between Afropavo and Pavo, and we were able to reject alternative phylogenetic hypotheses. Molecular clock estimates of the divergence time place the separation of Afropavo and Pavo in the late Miocene. We also discuss other relatives of Afropavo and Pavo and use this information to propose hypotheses regarding the evolution of ornamentation and sexual dimorphism within this group of pheasants.
Asunto(s)
Aves/genética , Animales , Aves/clasificación , Grupo Citocromo b/genética , ADN/análisis , Evolución Molecular , Filogenia , Alineación de SecuenciaRESUMEN
In the Neurospora Genome Project at the University of New Mexico, expressed sequence tags (ESTs) corresponding to three stages of the life cycle of the filamentous fungus Neurospora crassa are being analyzed. The results of a pilot project to identify expressed genes and determine their patterns of expression are presented. 1,865 partial complementary DNA (cDNA) sequences for 1,409 clones were determined using single-pass sequencing. Contig analysis allowed the identification of 838 unique ESTs and 156 ESTs present in multiple cDNA clones. For about 34% of the sequences, highly or moderately significant matches to sequences (of known and unknown function) in the NCBI database were detected. Approximately 56% of the ESTs showed no similarity to previously identified genes. Among genes with assigned function, about 43.3% were involved in metabolism, 32.9% in protein synthesis and 8.4% in RNA synthesis. Fewer were involved in defense (6%), cell signalling (3.4%), cell structure (3.4%) and cell division (2.6%).
Asunto(s)
Proteínas Fúngicas/genética , Genes Fúngicos , Neurospora crassa/genética , Neurospora crassa/fisiología , Secuencia de Aminoácidos , Secuencia de Bases , División Celular , ADN Complementario/química , Proteínas Fúngicas/biosíntesis , Expresión Génica , Biblioteca de Genes , Genoma Fúngico , Datos de Secuencia Molecular , Neurospora crassa/citología , ARN Mensajero/biosíntesisRESUMEN
The regulation of cellular growth and proliferation in response to environmental cues is critical for development and the maintenance of viability in all organisms. In unicellular organisms, such as the budding yeast Saccharomyces cerevisiae, growth and proliferation are regulated by nutrient availability. We have described changes in the pattern of protein synthesis during the growth of S. cerevisiae cells to stationary phase (E. K. Fuge, E. L. Braun, and M. Werner-Washburne, J. Bacteriol. 176:5802-5813, 1994) and noted a protein, which we designated Snz1p (p35), that shows increased synthesis after entry into stationary phase. We report here the identification of the SNZ1 gene, which encodes this protein. We detected increased SNZ1 mRNA accumulation almost 2 days after glucose exhaustion, significantly later than that of mRNAs encoded by other postexponential genes. SNZ1-related sequences were detected in phylogenetically diverse organisms by sequence comparisons and low-stringency hybridization. Multiple SNZ1-related sequences were detected in some organisms, including S. cerevisiae. Snz1p was found to be among the most evolutionarily conserved proteins currently identified, indicating that we have identified a novel, highly conserved protein involved in growth arrest in S. cerevisiae. The broad phylogenetic distribution, the regulation of the SNZ1 mRNA and protein in S. cerevisiae, and identification of a Snz protein modified during sporulation in the gram-positive bacterium Bacillus subtilis support the hypothesis that Snz proteins are part of an ancient response that occurs during nutrient limitation and growth arrest.
Asunto(s)
Proteínas Fúngicas/genética , Genes Fúngicos , Familia de Multigenes , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , División Celular , Clonación Molecular , Secuencia Conservada , Sondas de ADN , Proteínas Fúngicas/química , Genes Bacterianos , Genes de Plantas , Datos de Secuencia Molecular , Mutación , Fenotipo , ARN de Hongos/genética , ARN Mensajero/genéticaRESUMEN
Like other microorganisms, the yeast Saccharomyces cerevisiae responds to starvation by arresting growth and entering stationary phase. Because most microorganisms exist under conditions of nutrient limitation, the ability to tolerate starvation is critical for survival. Molecular analyses have identified changes in transcription, translation, and protein modification in stationary-phase cells. At the level of translation, the pattern of newly synthesized proteins in stationary-phase cells is surprisingly similar to the pattern of proteins synthesized during exponential growth. When limited for different nutrients, yeast strains may not enter stationary phase but opt for pathways such as pseudohyphal growth. If nutrient limitation continues, the end-point is likely to be a stationary-phase cell. Based on the results of recent studies, we propose a model for entry into stationary phase in which G(o) arrest is separable from acquisition of the ability to survive long periods of time without added nutrients.
Asunto(s)
Saccharomyces cerevisiae/citología , Carbono/metabolismo , División Celular , Regulación Fúngica de la Expresión Génica , Genes Fúngicos , Interfase , Modelos Biológicos , Biosíntesis de Proteínas , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMEN
We are interested in characterizing the process of entry into and the maintenance of the stationary phase. To identify proteins that are induced during growth to stationary phase, we examined protein synthesis in long-term stationary-phase cultures using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Although the total rate of protein synthesis declined when growth ceased after the postdiauxic phase, the pattern of proteins synthesized remained similar throughout the experimental period (28 days), except at the diauxic shift. At the diauxic shift most proteins detectable by 2D-PAGE undergo a transient reduction in their relative rate of synthesis that ends when cells resume growth during the postdiauxic phase. We conclude from this that the transient repression of protein synthesis at the diauxic shift is not directly associated with stationary-phase arrest. A number of proteins that are synthesized after exponential phase have been identified by 2D-PAGE. These proteins could be divided into three temporal classes depending upon when their synthesis became detectable. One postexponential protein, designated p35, was induced later than all other proteins, and its relative rate of synthesis increased throughout stationary phase. Unlike most postexponential proteins, p35 was not regulated by heat shock or glucose repression. We also observed that a direct correlation between steady-state mRNA accumulation and protein synthesis for another postexponential protein (Ssa3p) or two closely related constitutive proteins (Ssa1p and Ssa2p) did not exist. We conclude from this result that synthesis of proteins in stationary phase is regulated by mechanisms other than the control of steady-state mRNA accumulation.