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| Status |
Public on Sep 17, 2021 |
| Title |
Multiple in vivo roles for the C-terminal domain of the RNA chaperone Hfq |
| Organism |
Escherichia coli K-12 |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Hfq, a bacterial RNA chaperone, stabilizes small regulatory RNAs (sRNAs) and facilitates sRNA base-pairing with target mRNAs. Hfq has a conserved N-terminal domain and a poorly conserved disordered C-terminal domain (CTD). In a transcriptome-wide examination of the effects of a chromosomal CTD deletion (Hfq1-65), the Escherichia coli mutant was most defective for the accumulation of sRNAs that bind the proximal and distal faces of Hfq (Class II sRNAs), but other sRNAs also were affected. There were only modest effects on the levels of mRNAs, suggesting little disruption of sRNA-dependent regulation. However, cells expressing Hfq lacking the CTD deletion in combination with a weak distal face mutation were defective for the function of the Class II sRNA ChiX and repression of mutS, both dependent upon distal face RNA binding. Loss of the region between amino acids 66-72 was critical for this defect. The CTD region beyond amino acid 72 was not necessary for distal face-dependent regulation, but was needed for functions associated with the Hfq rim, seen most clearly in combination with a rim mutant. Our results suggest that the C-terminus collaborates in various ways with different binding faces of Hfq, leading to distinct outcomes for individual sRNAs.
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| Overall design |
Wild Type (WT) and Hfq65 (CTD deleted mutant) E.coli total RNA and RNA immunoprecipitated with Hfq were collected and sequenced with Illumina. The experiments were duplicated.
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| Contributor(s) |
Kavita K, Zhang A, Tai C, Majdalani N, Storz G, Gottesman S |
| Citation(s) |
35104863 |
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| Submission date |
Nov 05, 2019 |
| Last update date |
Mar 04, 2022 |
| Contact name |
Susan Gottesman |
| Organization name |
National Cancer Institute
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| Lab |
Lab of Molecular Biology
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| Street address |
37 Convent Drive
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
| GPL24570 |
Illumina HiSeq 2500 (Escherichia coli K-12) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA588270 |
| SRA |
SRP229111 |
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