'Drc', a structurally novel ssDNA-binding transcription regulator of N4-related bacterial viruses.
Amino Acid Sequence
Amino Acid Substitution
Binding Sites
Cloning, Molecular
DNA, Single-Stranded
/ chemistry
DNA, Viral
/ chemistry
DNA-Binding Proteins
/ chemistry
DNA-Directed RNA Polymerases
/ chemistry
Escherichia coli
/ genetics
Gene Expression
Genetic Vectors
/ chemistry
Models, Molecular
Nucleic Acid Conformation
Open Reading Frames
Promoter Regions, Genetic
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Protein Multimerization
Pseudomonas
/ virology
Pseudomonas Phages
/ genetics
Recombinant Proteins
/ chemistry
Transcription, Genetic
Viral Proteins
/ chemistry
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
10 01 2020
10 01 2020
Historique:
accepted:
23
10
2019
revised:
16
10
2019
received:
18
09
2019
pubmed:
15
11
2019
medline:
18
3
2020
entrez:
15
11
2019
Statut:
ppublish
Résumé
Bacterial viruses encode a vast number of ORFan genes that lack similarity to any other known proteins. Here, we present a 2.20 Å crystal structure of N4-related Pseudomonas virus LUZ7 ORFan gp14, and elucidate its function. We demonstrate that gp14, termed here as Drc (ssDNA-binding RNA Polymerase Cofactor), preferentially binds single-stranded DNA, yet contains a structural fold distinct from other ssDNA-binding proteins (SSBs). By comparison with other SSB folds and creation of truncation and amino acid substitution mutants, we provide the first evidence for the binding mechanism of this unique fold. From a biological perspective, Drc interacts with the phage-encoded RNA Polymerase complex (RNAPII), implying a functional role as an SSB required for the transition from early to middle gene transcription during phage infection. Similar to the coliphage N4 gp2 protein, Drc likely binds locally unwound middle promoters and recruits the phage RNA polymerase. However, unlike gp2, Drc does not seem to need an additional cofactor for promoter melting. A comparison among N4-related phage genera highlights the evolutionary diversity of SSB proteins in an otherwise conserved transcription regulation mechanism.
Identifiants
pubmed: 31724707
pii: 5625530
doi: 10.1093/nar/gkz1048
pmc: PMC7145618
doi:
Substances chimiques
DNA, Single-Stranded
0
DNA, Viral
0
DNA-Binding Proteins
0
Recombinant Proteins
0
Viral Proteins
0
DNA-Directed RNA Polymerases
EC 2.7.7.6
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
445-459Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
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