Regulation and RNA-binding properties of Hfq-like RNA chaperones in Bacillus anthracis

被引:12
|
作者
Panda, Gurudutta [1 ,2 ]
Tanwer, Pooja [1 ]
Ansari, Salim [1 ]
Khare, Devanshi [1 ]
Bhatnagar, Rakesh [1 ]
机构
[1] Jawaharlal Nehru Univ, Sch Biotechnol, New Delhi 110067, India
[2] Hamdard Univ, Dept Comp Sci, Bioinformat Sect, New Delhi 110062, India
来源
关键词
Hfq; RNA chaperones; Bacillus anthracis; Small RNAs; AbrB; CsfG; ESCHERICHIA-COLI HFQ; GRAM-POSITIVE BACTERIA; SM-LIKE PROTEINS; STAPHYLOCOCCUS-AUREUS; LISTERIA-MONOCYTOGENES; GENE-EXPRESSION; INTERACTION SURFACES; CRYSTAL-STRUCTURE; SUBTILIS; VIRULENCE;
D O I
10.1016/j.bbagen.2015.03.016
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: Small RNAs (sRNAs) are important modulators of gene expression in bacteria. Regulation by sRNAs is yet to be established in Bacillus anthracis. Here, regulation and RNA-binding properties of Hfq-like RNA chaperones in B. anthracis are investigated. Methods: Transcript levels were measured by RT-PCR. Proteins were cloned, purified, and their ability to bind sRNA was seen by EMSA. Regulators of Hfq1 were identified by in silico analysis and validated by EMSA. Conserved sRNAs were identified by homology search and their ability to bind Hfq1 was seen by EMSA. Residues crucial for sRNA binding were identified by mutational studies. Results: hfq1 and hfq3 showed expression during exponential phase on BHI medium, while hfq2 showed no expression. Hfq1 and Hfq3 formed hexamer and sRNA-Hfq complex, not Hfq2. In silica prediction and EMSA confirmed AbrB binding to the promoter of Hfq1. Homology search identified 7 sRNAs in B. anthracis. The sRNA CsfG showed binding to Hfq1 via residues Y24, F29, Q30, R32, K56, and H57. Conclusions: Hfq1 and Hfq3 can function as RNA chaperones in B. anthracis. The transition phase repressor AbrB might be responsible for the growth-dependent expression of Hfq1. The sporulation-specific sRNA CsfG binds to Hfq1 via its distal surface and requires an intact hexameric structure for forming CsfG-Hfq1 complex. General significance: This is the first report demonstrating the regulation and functional properties of Hfq-like RNA chaperones in B. anthracis and paves the path towards establishing sRNA-based regulation in B. anthracis. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:1661 / 1668
页数:8
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