Abstract:
UNLABELLED:Bacteriophage λ N protein binds boxB RNA hairpins in the nut (N utilization) sites of immediate early λ transcripts and interacts with host factors to suppress transcriptional termination at downstream terminators. In opposition to λ N, the Nun protein of HK022 binds the boxBs of coinfecting λ transcripts, interacts with a similar or identical set of host factors, and terminates transcription to suppress λ replication. Comparison of N-boxB and Nun-boxB nuclear magnetic resonance (NMR) structural models suggests similar interactions, though limited mutagenesis of Nun is available. Here, libraries of Nun's arginine-rich motif (ARM) were screened for the ability to exclude λ coinfection, and mutants were assayed for Nun termination with a boxB plasmid reporter system. Several Nun ARM residues appear to be immutable: Asp26, Arg28, Arg29, Arg32, Trp33, and Arg36. Asp26 and Trp33 appear to be unable to contact boxB and are not found at equivalent positions in λ N ARM. To understand if the requirement of Asp26, Trp33, and Arg36 indicated differences between HK022 Nun termination and λ N antitermination complexes, the same Nun libraries were fused to the activation domain of λ N and screened for clones able to complement N-deficient λ. Mutants were assayed for N antitermination. Surprisingly, Asp26 and Trp33 were still essential when Nun ARM was fused to N. Docking suggests that Nun ARM contacts a hydrophobic surface of the NusG carboxy-terminal domain containing residues necessary for Nun function. These findings indicate that Nun ARM relies on distinct contacts in its ternary complex and illustrate how protein-RNA recognition can evolve new regulatory functions. IMPORTANCE:λ N protein interacts with host factors to allow λ nut-containing transcripts to elongate past termination signals. A competing bacteriophage, HK022, expresses Nun protein, which causes termination of λ nut transcripts. λ N and HK022 Nun use similar arginine-rich motifs (ARMs) to bind the same boxB RNAs in nut transcripts. Screening libraries of Nun ARM mutants, both in HK022 Nun and in a λ N fusion, revealed amino acids essential to Nun that could bind one or more host factors. Docking suggests that NusG, which is present in both Nun termination and N antitermination, is a plausible partner. These findings could help understand how transcription elongation is regulated and illustrate how subtle differences allow ARMs to evolve new regulatory functions.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Tawk CS,Ghattas IR,Smith CAdoi
10.1128/JB.00466-15subject
Has Abstractpub_date
2015-11-01 00:00:00pages
3573-82issue
22eissn
0021-9193issn
1098-5530pii
JB.00466-15journal_volume
197pub_type
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.10.5713-5719.1989
更新日期:1989-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.126.1.198-204.1976
更新日期:1976-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.175.8.2197-2204.1993
更新日期:1993-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.145.1.654-656.1981
更新日期:1981-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.151.2.777-782.1982
更新日期:1982-08-01 00:00:00
abstract::The application of mitomycin C induction to 114 genetically diverse Streptococcus agalactiae strains generated 36 phage suspensions. On electron microscopy of the phage suspensions, it was possible to assign the phages to the Siphoviridae family, with three different morphotypes (A, B, and C). Phage genetic diversity ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00426-09
更新日期:2009-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.18.5820-5826.1992
更新日期:1992-09-01 00:00:00
abstract::Fowler, Audree V. (University of California, Los Angeles), and Irving Zabin. Effects of dimethylsulfoxide on the lactose operon in Escherichia coli. J. Bacteriol. 92:353-357. 1966.-Dimethylsulfoxide (DMSO) at a concentration of 5% (v/v) in the culture medium inhibits the growth of Escherichia coli to only a slight ext...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.92.2.353-357.1966
更新日期:1966-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2008-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.8.3459-3467.1988
更新日期:1988-08-01 00:00:00
abstract::A 12.1-kilobase PstI fragment from Rhizobium japonicum, which contains homology to both the Klebsiella pneumoniae and the Rhizobium meliloti nifH genes, was cloned into vector pHE3 . The nifH -homologous region was localized on the restriction enzyme cleavage map by Southern blot hybridization experiments. DNA fragmen...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.158.3.1005-1011.1984
更新日期:1984-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.130.3.1253-1261.1977
更新日期:1977-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01623-14
更新日期:2014-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.24.6484-6492.1998
更新日期:1998-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.5.2063-2069.1988
更新日期:1988-05-01 00:00:00
abstract::Mushin, Rose (University of Melbourne, Victoria, Australia) and Frances M. Ashburner. Gastrointestinal microflora of mutton birds (Puffinus tenuirostris) in relation to "limy" disease. J. Bacteriol. 83:1260-1267. 1962.-The aerobic bacterial intestinal flora of mutton birds, healthy and affected with "limy" disease, wa...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.83.6.1260-1267.1962
更新日期:1962-06-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of bacteriology
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doi:10.1128/JB.121.2.416-421.1975
更新日期:1975-02-01 00:00:00
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更新日期:2005-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JB.180.8.2160-2166.1998
更新日期:1998-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2012-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2019-09-06 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.183.23.6740-6745.2001
更新日期:2001-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.173.19.6162-6167.1991
更新日期:1991-10-01 00:00:00
abstract::The effect of motility on the competitive success of Rhizobium meliloti in nodule production was investigated. A motile strain formed more nodules than expected when mixed at various unfavorable ratios with either flagellated or nonflagellated nonmotile derivatives. We conclude that motility confers a selective advant...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.148.2.728-729.1981
更新日期:1981-11-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JB.113.2.1034-1045.1973
更新日期:1973-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.116.2.757-763.1973
更新日期:1973-11-01 00:00:00
abstract::Previously we have shown that flagella and the O-specific polysaccharide of lipopolysaccharide play a role in colonization of the potato root by plant growth-promoting Pseudomonas strains WCS374 and WCS358. In this paper, we describe a novel cell surface-exposed structure in Pseudomonas putida WCS358 examined with a s...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.178.7.1955-1961.1996
更新日期:1996-04-01 00:00:00
abstract::Buck, C. A. (Kansas State University, Manhattan), R. L. Anacker, F. S. Newman, and A. Eisenstark. Phage isolated from lysogenic Bacillus anthracis. J. Bacteriol. 85:1423-1430. 1963.-Bacillus anthracis cultures (a total of 311) were obtained from several laboratories throughout the world; 139 of these were screened for...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.85.6.1423-1430.1963
更新日期:1963-06-01 00:00:00