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Published on: February 9, 2015
Host factor Hfq of Escherichia coli stimulates elongation of poly(A) tails by poly(A) polymerase I
1Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, 75005 Paris, France.
Abstract:
Current evidence suggests that the length of poly(A) tails of bacterial mRNAs result from a competition between poly(A) polymerase and exoribonucleases that attack the 3' ends of RNAs. Here, we show that host factor Hfq is also involved in poly(A) tail metabolism. Inactivation of the hfq gene reduces the length of poly(A) tails synthesized at the 3' end of the rpsO mRNA by poly(A) polymerase I in vivo. In vitro, Hfq stimulates synthesis of long tails by poly(A) polymerase I. The strong binding of Hfq to oligoadenylated RNA probably explains why it stimulates elongation of primers that already harbor tails of 20-35 A. Polyadenylation becomes processive in the presence of Hfq. The similar properties of Hfq and the PABPII poly(A) binding protein, which stimulates poly(A) tail elongation in mammals, indicates that similar mechanisms control poly(A) tail synthesis in prokaryotes and eukaryotes.
Insights
Host factor Hfq regulates bacterial mRNA poly(A) tail length by stimulating poly(A) polymerase I. This finding reveals a conserved mechanism for polyadenylation in prokaryotes and eukaryotes.
Area of Science:
- Molecular Biology
- Bacterial Genetics
- RNA Metabolism
Background:
- Bacterial mRNA poly(A) tail length is determined by a balance between polyadenylation and degradation.
- Host factor Hfq's role in poly(A) tail metabolism was previously uncharacterized.
Purpose of the Study:
- To investigate the involvement of host factor Hfq in bacterial mRNA poly(A) tail metabolism.
- To elucidate the mechanism by which Hfq influences polyadenylation.
Main Methods:
- In vivo studies involving inactivation of the hfq gene and analysis of rpsO mRNA poly(A) tail length.
- In vitro experiments using poly(A) polymerase I and Hfq to assess polyadenylation activity.
- Investigating the binding properties of Hfq to oligoadenylated RNA.
Main Results:
- Inactivation of the hfq gene led to shorter poly(A) tails on rpsO mRNA.
- Hfq significantly stimulated the synthesis of long poly(A) tails by poly(A) polymerase I in vitro.
- Hfq demonstrated strong binding to RNA with existing poly(A) tails (20-35 nucleotides), promoting processive polyadenylation.
Conclusions:
- Host factor Hfq plays a crucial role in regulating bacterial mRNA poly(A) tail length.
- Hfq enhances poly(A) tail elongation by stimulating poly(A) polymerase I activity.
- The findings suggest conserved mechanisms for poly(A) tail synthesis control in both prokaryotes and eukaryotes, similar to mammalian poly(A) binding protein II (PABPII).
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