This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kaur, P Articles by Rosen, B P Search for Related Content PubMed PubMed Citation Articles by Kaur, P Articles by Rosen, B P

research-article

Complementation between nucleotide binding domains in an anion-translocating ATPase.

Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201.

ABSTRACT

The catalytic component of the oxyanion-translocating ATPase of the plasmid-encoded ars operon of Escherichia coli is a homodimer of the ArsA protein. This enzyme is an oxyanion-stimulated ATPase with two consensus nucleotide binding sequences in each subunit, one in the N-terminal (A1) half and one in the C-terminal (A2) half of the ArsA protein. The two halves of both the arsA gene and the ArsA protein exhibit similar nucleotide and amino acid sequences, respectively. The two halves of the arsA gene were subcloned into compatible plasmids. Neither alone was sufficient to confer resistance, but cells in which the arsA1 and arsA2 half genes were coexpressed were resistant to arsenicals. Genetic complementation was also observed in cells bearing plasmids with point mutations in the two halves of the arsA gene and between cells with plasmids carrying combinations of the arsA1 or arsA2 subclones and point mutations. In every case, complementation was observed only when one plasmid contained a wild-type arsA1 sequence and the other contained a wild-type arsA2 sequence. These results demonstrate that both sites are required for resistance but that the two nucleotide binding domains need not reside in a single polypeptide. We propose a model in which the ArsA dimer has two catalytic units, each composed of an A1 domain from one monomer and an A2 domain from the other monomer.

This article has been cited by other articles:

• Kaur, P. (1999). The Anion-stimulated ATPase ArsA Shows Unisite and Multisite Catalytic Activity. J. Biol. Chem. 274: 25849-25854 [Abstract] [Full Text]  
• Walmsley, A. R., Zhou, T., Borges-Walmsley, M. I., Rosen, B. P. (1999). The ATPase Mechanism of ArsA, the Catalytic Subunit of the Arsenite Pump. J. Biol. Chem. 274: 16153-16161 [Abstract] [Full Text]  
• Kaur, P., Russell, J. (1998). Biochemical Coupling between the DrrA and DrrB Proteins of the Doxorubicin Efflux Pump of Streptomyces peucetius. J. Biol. Chem. 273: 17933-17939 [Abstract] [Full Text]  
• Ramaswamy, S., Kaur, P. (1998). Nucleotide Binding to the C-terminal Nucleotide Binding Domain of ArsA. STUDIES WITH AN ATP ANALOGUE, 5'-p-FLUOROSULFONYLBENZOYLADENOSINE (FSBA). J. Biol. Chem. 273: 9243-9248 [Abstract] [Full Text]  
• Li, J., Rosen, B. P. (1998). Steric Limitations in the Interaction of the ATP Binding Domains of the ArsA ATPase. J. Biol. Chem. 273: 6796-6800 [Abstract] [Full Text]  
• Zhou, T., Rosen, B. P. (1997). Tryptophan Fluorescence Reports Nucleotide-induced Conformational Changes in a Domain of the ArsA ATPase. J. Biol. Chem. 272: 19731-19737 [Abstract] [Full Text]  
• Li, J., Liu, S., Rosen, B. P. (1996). Interaction of ATP Binding Sites in the ArsA ATPase, the Catalytic Subunit of the Ars Pump. J. Biol. Chem. 271: 25247-25252 [Abstract] [Full Text]  
• Jia, H., Kaur, P. (2001). Role of the Linker Region of the Anion-stimulated ATPase ArsA. EFFECT OF DELETION AND POINT MUTATIONS IN THE LINKER REGION. J. Biol. Chem. 276: 29582-29587 [Abstract] [Full Text]