Cloning and nucleotide sequence analysis of the Streptococcus mutans membrane-bound, proton-translocating ATPase operon.
The function of the membrane-bound ATPase in S. mutans is to regulate cytoplasmic pH values for the purpose of maintaining delta pH. Previous studies have shown that as part of its acid-adaptive ability, S. mutans is able to increase H(+)-ATPase levels in response to acidification. As part of the study of ATPase regulation in S. mutans, we have cloned the ATPase operon and determined its genetic organization. The structural genes from S. mutans were found to be in the order: c, a, b, delta, alpha, gamma, beta, and epsilon; where c and a were reversed from the more typical bacterial organization. The operon contained no I gene homologue but was preceded by a 239-bp intergenic space. Deduced aa sequences from open reading frames indicated that genes encoding homologues of glycogen phosphorylase and nonphosphorylating, NADP-dependent glyceraldehyde-3-phosphate dehydrogenase flank the H(+)-ATPase operon, 5' and 3' respectively. Sequence analysis indicated the presence of three inverted-repeat nt sequences in the glgP-uncE intergenic space. Primer extension analysis of mRNAs prepared from batch-grown or steady-state cultures demonstrated that the transcriptional start site did not change as a function of culture pH value. The data suggest that potential stem-and-loop structures in the promoter region of the operon do not function to alter the starting position of ATPase-specific mRNA transcription.
Smith
A J
AJ
Department of Dental Research, School of Medicine and Dentistry, University of Rochester, NY 14642, USA.
Quivey
R G
RG
Faustoferri
R C
RC
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D025243:Bacterial Proton-Translocating ATPases; D002462:Cell Membrane / Q000201:enzymology; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial / Q000235:genetics*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D009876:Operon / Q000235:genetics*; D006005:Phosphorylases / Q000235:genetics; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013295:Streptococcus mutans / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; C074429:atpE protein, E coli; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D006005:Phosphorylases; D025243:Bacterial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases