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RNA Binding by the Campylobacter jejuni Post-transcriptional Regulator CsrA
is a Gram-negative rod-shaped bacterium that commensally inhabits the intestinal tracts of livestock and birds, and which also persists in surface waters. is a leading cause of foodborne gastroenteritis, and these infections are sometimes associated with the development of post-infection sequelae su...
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Published in: | Frontiers in microbiology 2019-08, Vol.10, p.1776-1776 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | is a Gram-negative rod-shaped bacterium that commensally inhabits the intestinal tracts of livestock and birds, and which also persists in surface waters.
is a leading cause of foodborne gastroenteritis, and these infections are sometimes associated with the development of post-infection sequelae such as Guillain-Barré Syndrome. Flagella are considered a primary virulence factor in
as these organelles are required for pathogenicity-related phenotypes including motility, biofilm formation, host cell interactions, and host colonization. The post-transcriptional regulator CsrA regulates the expression of the major flagellin FlaA by binding to
mRNA and repressing its translation. Additionally, CsrA has previously been shown to regulate 120-150 proteins involved in diverse cellular processes. The amino acid sequence of
CsrA is significantly different from that of
CsrA, and no previous research has defined the amino acids of
CsrA that are critical for RNA binding. In this study, we used
SELEX to identify the consensus RNA sequence mAwGGAs to which
CsrA binds with high affinity. We performed saturating site-directed mutagenesis on
CsrA and assessed the regulatory activity of these mutant proteins, using a reporter system encoding the 5' untranslated region (5' UTR) upstream of
linked translationally to the
gene. These assays allowed us to identify 19 amino acids that were involved in RNA binding by CsrA, with many but not all of these amino acids clustered in predicted beta strands that are involved in RNA binding by
CsrA. Decreased
mRNA binding by mutant CsrA proteins L2A and A36V was confirmed by electrophoretic mobility shift assays. The majority of the amino acids implicated in RNA binding were conserved among diverse
species. |
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ISSN: | 1664-302X 1664-302X |
DOI: | 10.3389/fmicb.2019.01776 |