Abstract
The Staphylococcus aureus chromosomal gene pcrA, identified by mutations, such as pcrA3, that affect plasmid pT181 replication, has been cloned and sequenced. The pcrA gene encodes a protein with significant similarity (40% identity) to two Escherichia coli helicases: the helicase II encoded by the uvrD gene and the Rep helicase. The pcrA3 mutation was found to be a C to T transition leading to a threonine to isoleucine substitution at amino acid residue 61 of the protein. The pcrA gene seems to belong to an operon containing at least one other gene, tentatively named pcrB, upstream from pcrA. The PcrA protein was shown to be essential for cell viability and overproduction has deleterious effects on the host and plasmid replication.
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References
Allen GC, Kornberg A (1991) Fine balance in the regulation of DnaB helicase by DnaC protein in replication in Escherichia coli. J Biol Chem 256:5287–5293
Arai N, Arai K-I, Kornberg A (1981) Complex of Rep protein with ATP and DNA as basis for helicase action. J Biol Chem 256:5287–5293
Carleton S, Projan SJ, Highlander SK, Moghazeh S, Novick RP (1984) Control of pT181 replication. II. Mutational analysis. EMBO J 3:2407–2414
Cerretti DP, Dean D, Davis GR, Bedwell DM, Nomura M (1983) The spc ribosomal protein operon of Escherichia coli: sequence and cotranscription of the ribosomal protein genes and a protein export gene. Nucleic Acids Res 11:2599–2616
Colasanti J, Denhardt DT (1987) The Escherichia coli rep mutation. X. Consequences of increased and decreased Rep protein levels. Mol Gen Genet 209:382–390
Finch PW, Emmerson PT (1984) The nucleotide sequence of the uvrD gene of E. coli. Nucleic Acids Res 12:5789–5799
Gilchrist CA, Denhardt DT (1987) Escherichia coli rep gene: sequence of the gene, the encoded helicase, and its homology with uvrD. Nucleic Acids Res 15:465–475
Götz F, Ahrne S, Lindberg M (1981) Plasmid transfer and genetic recombination by protoplast fusion in staphylococci. J Bacteriol 145:74–81
Hodgman TC (1988) A new superfamily of replicative proteins. Nature 333:22–23
Iordanescu S, Bargonetti J (1989) Staphylococcus aureus chromosomal mutations that decrease efficiency of Rep utilization in replication of pT181 and related plasmids. J Bacteriol 171:4501–4503
Iordanescu S, Basheer R (1991) The Staphylococcus aureus mutation pcrA3 leads to the accumulation of pT181 replication initiation complexes. J Mol Biol 221:1183–1189
Iordanescu S, Surdeanu M (1980) Complementation of a plasmid replication defect by autonomous, incompatible plasmids in Staphylococcus aureus. Plasmid 4:1–7
Jones C, Khan SA (1986) Nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus. J Bacteriol 166:29–33
Kumura K, Sekiguchi M (1984) Identification of the uvrD gene product of Escherichia coli as DNA helicase II and its induction by DNA-damaging agents. J Biol Chem 259:1560–1565
Lohman TM, Chao K, Green JM, Sage S, Runyon GT (1989) Large-scale purification and characterization of the Escherichia coli rep gene product. J Biol Chem 264:10139–10147
Maples VF, Kushner SR (1982) DNA repair in Escherichia coli: identification of the uvrD gene product. Proc Natl Acad Sci USA 79:5616–5620
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:10–89
Nesin M, Svec P, Lupski JR, Godson GN, Kreiswirth B, Kornblum J, Projan SJ (1990) Cloning and nucleotide sequence of a chromosomally encoded tetracycline resistance determinant, tetA (M), from a pathogenic, methicillin-resistant strain of Staphylococcus aureus. Antimicrob Agents Chemother 34:2273–2276
Novick RP, Richmond MH (1965) Nature and interactions of the genetic elements governing penicillinase synthesis in Staphylococcus aureus. J Bacteriol 90:467–480
Novick RP, Adler GK, Projan SJ, Carleton S, Highlander S, Gruss A, Khan SA, Iordanescu S (1984) Control of pT181 replication. I. The pT181 copy control function acts by inhibiting the synthesis of a replication protein. EMBO J 3:2399–2405
Novick RP, Edelman I, Löfdahl S (1986) Small Staphylococcus aureus plasmids are transduced as linear multimers that are formed and resolved by replicative processes. J Mol Biol 192:209–220
Novick RP, Iordanescu S, Projan SJ, Kornblum J, Edelman I (1989) pT181 plasmid replication is regulated by a countertranscriptdriven transcriptional attenuator. Cell 59:395–404
O'Callaghan CH, Morris A, Kirby SM, Shlager AH (1972) Novel method for detection of β-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother 1:283–288
Oeda K, Horiuchi T, Sekiguchi M (1982) The uvrD gene of E. coli encodes a DNA-dependent ATPase. Nature 298:98–100
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Projan SJ, Kornblum J, Moghazeh SL, Edelman I, Gennaro ML, Novick RP (1985) Comparative sequence and functional analysis of pT181 and pC221, cognate plasmid replicons from Staphylococcus aureus. Mol Gen Genet 199:452–464
Runyon GT, Lohman TM (1989) Escherichia coli helicase II (UvrD) protein can completely unwind fully duplex linear and nicked circular DNA. J Biol Chem 264:17502–17512
Sambrook J, Fritsch EF, Maniatis F (1989) Molecular cloning: A Laboratory manual. Cold Spring Laboratory Press, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 85:5463–5467
Taucher-Scholz G, Abdel-Monem M, Hoffmann-Berling H (1983) Functions of DNA helicases in Escherichia coli. pp. 65–76. In Mechanisms of DNA Replication and Recombination, Alan R. Liss, Inc., New York
Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the α- and β-subunits of ATP synthase, myosin kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951
Washburn BK, Kushner SR (1991) Construction and analysis of deletions in the structural gene (uvrD) for helicase II of Escherichia coli. J Bacteriol 178:2569–2575
Youngman P (1987) Plasmid vectors for recovering and exploiting Tn917 transpositions in Bacillus and other Gram-positive bacteria. In: Hardy KG (ed.) Plasmids — a practical approach. IRL Press, Oxford, pp 79–104
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Iordanescu, S. Characterization of the Staphylococcus aureus chromosomal gene pcrA, identified by mutations affecting plasmid pT181 replication. Molec. Gen. Genet. 241, 185–192 (1993). https://doi.org/10.1007/BF00280216
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DOI: https://doi.org/10.1007/BF00280216