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J Bacteriol, April 1998, p. 1929-1938, Vol. 180, No. 7
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Escherichia coli mrsC Is an Allele of hflB, Encoding a Membrane-Associated ATPase and Protease That Is Required for mRNA Decay

Rong-fu Wang,dagger Eileen B. O'Hara, Marti Aldea,Dagger Cornelia I. Bargmann,§ Heather Gromley, and Sidney R. Kushner*

Department of Genetics, University of Georgia, Athens, Georgia 30602-7223

Received 29 September 1997/Accepted 28 January 1998

The mrsC gene of Escherichia coli is required for mRNA turnover and cell growth, and strains containing the temperature-sensitive mrsC505 allele have longer half-lives than wild-type controls for total pulse-labeled and individual mRNAs (L. L. Granger et al., J. Bacteriol. 180:1920-1928, 1998). The cloned mrsC gene contains a long open reading frame beginning at an initiator UUG codon, confirmed by N-terminal amino acid sequencing, encoding a 70,996-Da protein with a consensus ATP-binding domain. mrsC is identical to the independently identified ftsH gene except for three additional amino acids at the N terminus (T. Tomoyasu et al., J. Bacteriol. 175:1344-1351, 1993). The purified protein had a Km of 28 µM for ATP and a Vmax of 21.2 nmol/µg/min. An amino-terminal glutathione S-transferase-MrsC fusion protein retained ATPase activity but was not biologically active. A glutamic acid replacement of the highly conserved lysine within the ATP-binding motif (mrsC201) abolished the complementation of the mrsC505 mutation, confirming that the ATPase activity is required for MrsC function in vivo. In addition, the mrsC505 allele conferred a temperature-sensitive HflB phenotype, while the hflB29 mutation promoted mRNA stability at both 30 and 44°C, suggesting that the inviability associated with the mrsC505 allele is not related to the defect in mRNA decay. The data presented provide the first direct evidence for the involvement of a membrane-bound protein in mRNA decay in E. coli.


* Corresponding author. Mailing address: Department of Genetics, Life Sciences Building, University of Georgia, Athens, GA 30602-7223. Phone: (706) 542-8000. Fax: (706) 542-3910. E-mail: skushner{at}uga.cc.uga.edu.

dagger Present address: Surgery Branch, National Cancer Institute, Bethesda, MD 20892.

Dagger Present address: Departament de Ciències Mèdiques Bàsques, Universitat de Lleida, 25196 Lleida, Spain.

§ Present address: Department of Anatomy, University of California, San Francisco, CA 94143-0452.




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