Identification of amino acid residues in the catalytic domain of RNase E essential for survival of Escherichia coli: Functional analysis of DNase I subdomainopen access
- Authors
- Shin, Eunkyoung; Go, Hayoung; Yeom, Ji-Hyun; Won, Miae; Bae, Jeehyeon; Han, Seung Hyun; Han, Kook; Lee, Younghoon; Ha, Nam-Chul; Moore ,Christopher J.; Sohlberg, Bjoern; Cohen, Stanley N.; Lee, Kangseok
- Issue Date
- Aug-2008
- Publisher
- GENETICS SOC AM
- Citation
- GENETICS, v.179, no.4, pp 1871 - 1879
- Pages
- 9
- Journal Title
- GENETICS
- Volume
- 179
- Number
- 4
- Start Page
- 1871
- End Page
- 1879
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23675
- DOI
- 10.1534/genetics.108.088492
- ISSN
- 0016-6731
1943-2631
- Abstract
- RNase E is all essential Escherichia coli endoribonuclease that plays a major endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell. To better understand the molecular mechanisms of RNase E action, we performed a genetic screen for amino acid substitution in the catalytic domain of the protein (N-Rne) that knock down the ability of RNase E to support survival of E. coli. Comparative phylogenetic analysis of RNase E homologs shows that wild-type residues at these mutated positions are nearly invariably conserved. Cells conditionally expressing these N-Rne mutants in the absence of wild-type RNase E show a decrease in copy number of plasmids regulated by the RNase E substrate RNA 1, and accumulation of 5S ribosomal RNA, M1 RNA, and tRNA(Asn) precursors, as has been found in Rne-depleted cells, suggesting that the inability of these mutants to support cellular growth results from loss of ribonucleolytic activity. Purified mutant proteins containing an amino acid substitution in the DNase 1 subdomain, which is spatially distant from the catalytic site posited from crystallographic studies, showed defecting binding to an RNase E substrate, p23 RNA, but still retained RNA cleavage activity-implicating a previously unidentified structural motif in the DNase 1 subdomain in the binding of RNase E to targeted RNA molecules, demonstrating the role of the DNase 1 domain in RNase E activity. subdoinain, which is spatially distant from the Catalytic site posited from ctystallographic studies, showed
- Files in This Item
-
- Appears in
Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
- College of Pharmacy > School of Pharmacy > 1. Journal Articles
![qrcode](https://api.qrserver.com/v1/create-qr-code/?size=55x55&data=https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23675)
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.