Programmable Acetylation Modification of Bacterial Proteins by a Cas12a-Guided Acetyltransferase.

Protein lysine acetylation (PLA) is a crucial post-translational modification in organisms that regulates a variety of metabolic and physiological activities. Many advances have been made in PLA-related research; however, the quick and accurate identification of causal relationships between specific protein acetylation events and phenotypic outcomes at the proteome level remains challenging due to the lack of efficient targeted modification techniques. In this study, based on the characteristics of transcription-translation coupling in bacteria, we designed and constructed an in situ targeted protein acetylation (TPA) system integrating the dCas12a protein, guiding element crRNA, and bacterial acetylase At2. Rapid identification of multiple independent protein acetylation and cell phenotypic analyses in Gram-negative Escherichia coli and Gram-positive Clostridium ljungdahlii demonstrated that TPA is a specific and efficient targeting tool for protein modification studies and engineering.

Fungal CFEM effectors negatively regulate a maize wall-associated kinase by interacting with its alternatively spliced variant to dampen resistance.

The fungus Fusarium graminearum causes a devastating disease Gibberella stalk rot of maize. Our knowledge of molecular interactions between F. graminearum effectors and maize immunity factors is lacking. Here, we show that a group of cysteine-rich common in fungal extracellular membrane (CFEM) domain proteins of F. graminearum are required for full virulence in maize stalk infection and that they interact with two secreted maize proteins, ZmLRR5 and ZmWAK17ET. ZmWAK17ET is an alternative splicing isoform of a wall-associated kinase ZmWAK17. Both ZmLRR5 and ZmWAK17ET interact with the extracellular domain of ZmWAK17. Transgenic maize overexpressing ZmWAK17 shows increased resistance to F. graminearum, while ZmWAK17 mutants exhibit enhanced susceptibility to F. graminearum. Transient expression of ZmWAK17 in Nicotiana benthamiana triggers hypersensitive cell death, whereas co-expression of CFEMs with ZmWAK17ET or ZmLRR5 suppresses the ZmWAK17-triggered cell death. Our results show that ZmWAK17 mediates stalk rot resistance and that F. graminearum delivers apoplastic CFEMs to compromise ZmWAK17-mediated resistance.

The complete mitochondrial genome of the Daimio tethys (Lepidoptera: Hesperoidea: Hesperiidae).

The complete mitochondrial genome of Daimio tethys (Hesperoidea: Hesperiidae) is a circular molecule of 15,341 bp in length, containing 37 typical animal mitochondrial genes: 13 protein coding genes (ATP6, ATP8, COI-III, ND1-6, ND4L, Cytb), 2 rRNA genes, 22 tRNA genes and a non-coding AT-rich region. Its gene order and content are identical to all other available butterfly mitogenomes. All PCGs initiate with typical ATN codons, except for COI, which is initiated by the CGA codon as observed in other butterfly species. Ten PCGs use complete termination codons TAA or TAG, whereas the COI, COII and ND5 genes end with a single T. A total of 179 bp of intergenic spacers are interspersed in 11 regions, ranging in size from 1 to 82 bp. The AT-rich region is 415 bp long and contains some conserved structures characteristic of the lepidopteran mitogenomes, including the motif ATAGA followed by a 17 bp poly-T stretch and a microsatellite-like (AT)8 element preceded by the ATTA motif.

The complete mitochondrial genome of Ideopsis similis (Lepidoptera: Nymphalidae: Danainae).

The complete mitochondrial nucleotide sequence of Ideopsis similis is 15,200 bp in size, containing typical metazoan 37 genes: 13 protein-coding genes (PCGs), 22 tRNA genes, 2 ribosomal RNA genes and 1 non-coding AT-rich region. All the 13 PCGs initiated with ATN, except for COI gene which started by CGA codon. Nine PCGs used the complete termination codon (TAA), whereas the COI, COII, ND4 and ND5 genes ended with single T. The two rRNA genes (rrnL and rrnS) were 1318 bp and 786 bp in size, with their AT contents of 84.7% and 85.7%, respectively. All tRNA genes display typical secondary cloverleaf structures except for trnS1(AGN) which loses DHU arm. The 404 bp long AT-rich region contains several features common to the other lepidopterans, such as the ATAGA motif followed by a 19 bp poly-T stretch, a 9 bp poly-T stretch and some microsatellite-like (AT)n elements.

The complete mitochondrial genome of Triphysa phryne (Lepidoptera: Nymphalidae: Satyrinae).

The complete mitochondrial genome (mitogenome) sequence of Triphysa phryne (Lepidoptera: Nymphalidae: Satyrinae) was determined in this study. The mitogenome is 15,143 bp in length, containing 37 typical animal mitochondrial genes: 13 putative protein-coding genes (PCGs), 2 ribosomal RNAs, 22 transfer RNAs and a non-coding AT-rich region. Its gene content and order are identical to those of other lepidopteran mitogenomes. All protein-coding genes (PCGs) are initiated by ATN codons, except for COI gene which uses CGA as its start codon. Nine PCGs terminate in the common stop TAA, whereas the COI, COII, ND5 and ND4 genes end with single T. All tRNA genes showed typical secondary cloverleaf structures except for the tRNA(Ser)(AGN), which has a simple loop with the absence of its DHU stem. The 316 bp AT-rich region contains several features common to the other lepidopterans, such as the motif ATAGA followed by an 19-bp poly-T stretch and two microsatellite-like (TA)8(AT) and (TA)4 elements preceded by the ATTTA motif.

The complete mitochondrial genome of the Byasa alcinous (Lepidoptera: Papilionidae: Papilioninae).

The complete mitochondrial genome (mitogenome) of Byasa alcinous (Lepidoptera: Papilionidae: Papilioninae) is a circular molecule of 15,266 bp in length, containing 37 typical insect mitochondrial genes: 13 protein coding genes (PCGs), 2 rRNA genes, 22 tRNA genes and a non-coding AT-rich region. Its gene order and arrangement are identical to all other available butterfly mitogenomes. All PCGs start with a typical ATN initiation codon, except for COI, which is initiated by the CGA codon as observed in other butterfly species. Ten PCGs terminate in the complete stop codon TAA or TAG, whereas the COI, COII and ND4 genes end with single T. Ten intergenic spacers (73 bp in total), and 12 overlapping regions (28 bp in total) are dispersed throughout the whole genome. The non-coding AT-rich region is 405 bp long and contains some conserved structures similar to those found in other butterfly mitogenomes, such as the motif ATAGA followed by a 12-bp poly-T stretch and a microsatellite-like (AT)14 element preceded by the ATTTA motif. Additionally, a 11-bp poly-T sequences and a microsatellite-like (AT)7 repeated elements are detected in this region.

The complete mitochondrial genome of Tirumala limniace (Lepidoptera: Nymphalidae: Danainae).

The complete mitochondrial nucleotide sequence of Tirumala limniace is 15,285 bp in size, containing typical metazoan 37 genes: 13 protein-coding genes (PCGs), 22tRNA genes, 2 ribosomal RNA genes and one noncoding AT-rich region. All the 13 PCGs initiated with ATN, except for COI gene which is started by CGA codon. Nine PCGs use the complete termination codon (TAA), whereas the COI, COII, ND4 and ND5 genes end with single T. A total of 132 bp intergenic spacers and a total of 37 bp overlapping sequences are interspersed throughout the whole genome. The two rRNA genes (rrnL and rrnS) are 1352 bp and 779 bp in size, with their AT contents of 84.6% and 85.5%, respectively. All tRNA genes display typical secondary cloverleaf structures except for trnS1 (AGN) which loses DHU arm. The 438 bp long AT-rich region contains several features common to the other lepidopterans, such as the ATAGA motif followed by a 18 bp poly-T stretch, a 9 bp poly-T stretch, a 10 bp poly-A stretch and some microsatellite-like (AT)n elements.