Cloning and primary structure of Staphylococcus aureus DNA topoisomerase IV: a primary target of fluoroquinolones

Molecular Microbiology - Tập 13 Số 4 - Trang 641-653 - 1994
Lucía Ferrero1, Béatrice Cameron1, B Manse1, D Lagneaux1, J Crouzet1, A Famechon2, Francis Blanche2
1Service de Biologie Moléculaire, Département des Biotechnologies
2Département Analyse, Rhône-Poulenc Rorer S.A., Centre de Recherche de Vitry-Alfortville, 13 Quai Jules Guesdes B.P. 14, 94403 Vitry-sur-Seine Cédex, France.

Tóm tắt

SummaryA 4.6 kb Staphylococcus aureus DNA fragment containing DNA gyrase‐like genes (grlA and grlB) was cloned and sequenced. The proteins GrlA and GrlB exhibit more than 30% identity with E. coli DNA topoisomerase IV sub units and with the gyrase subunits from S. aureus and Escherichia coli. The combined E. coli cell extracts of GrlA and GrlB overproducing strains catalysed ATP‐dependent relaxation and decatenation specific to DNA topoisomerase IV. The temperature‐sensitive phenotype of Salmonella typhimurium parC and parE mutants was complemented by the S. aureus grlA and grlB genes, when the two genes were co‐expressed. These results show that GrlA and GrlB are the subunits of S. aureus DNA topoisomerase IV. The GyrA subunit of DNA gyrase has been previously defined as a primary target of quinolones based on genetic and biochemical experiments essentially carried out in E. coli. Single‐point mutations occurring in the‘quinolone resistance‐determining region’(QRDR) of GyrA were found in bacteria exhibiting quinolone resistance, the most common mutation being a substitution of Ser‐83 on the E. coli GyrA sequence. We analysed eight S. aureus fluoroquinolone‐resistant clinical isolates and observed that mutations in the QRDR of GyrA are not present in the low‐quinolone‐resistant isolates. In contrast, Ser‐80 of GrlA, which corresponds to Ser‐83 of E. coli GyrA, is substituted to Phe or Tyr in both high‐ and low‐quinolone‐resistant isolates. We propose that DNA topoisomerase IV is a primary target of fluoroquinolones in S. aureus.

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Tài liệu tham khảo

10.1093/nar/15.2.771

10.1016/0092-8674(92)90356-H

10.1016/0378-1119(81)90080-9

10.1126/science.259.5092.163-a

10.1111/j.1365-2958.1990.tb00687.x

10.1128/AAC.33.6.886

10.1016/0378-1119(87)90158-2

Dessen P.C., 1990, Bisance: a french service for the access to biomolecular sequence databases, Comp Appl Biosciences, 6, 355

10.1128/AAC.36.5.1166

10.1016/0378-1119(90)90148-K

10.1016/0378-1119(88)90330-7

10.1128/jb.173.2.642-648.1991

Hooper D.C., 1993, Quinolone Antimicrobial Agents, 53

Horowitz D.S., 1987, Mapping of the active site tyrosine of Escherichia coli DNA gyrase, J Biol Chem, 262, 5339, 10.1016/S0021-9258(18)61193-7

Huang W.M., 1992, Molecular Biology of DNA Topoisomerases and its Application to Chemotherapy, 39

10.1093/nar/12.1Part1.203

10.1016/0092-8674(90)90172-B

Kato J., 1992, Purification and characterization of DNA topoisomerase‐IV in Escherichia coli, J Biol Chem, 267, 25676, 10.1016/S0021-9258(18)35660-6

10.1016/0092-8674(87)90503-4

10.1038/305709a0

10.1073/pnas.80.3.697

Luttinger A.L., 1991, A cluster of genes that affects nucleoid segregation in Salmonella typhimurium, New Biol, 3, 687

10.1128/jb.174.5.1596-1603.1992

10.1093/jac/30.4.409

10.1093/nar/13.7.2251

10.1126/science.257.5073.1064

10.1146/annurev.ge.17.120183.002435

Novick R.P., 1990, Molecular Biology of the Staphylococci, 1

10.1093/nar/18.19.5880

10.1016/S0021-9258(20)80551-1

10.1073/pnas.90.18.8571

10.3109/10409239109114072

Sambrook J., 1989, Molecular Cloning: A Laboratory Manual

10.1128/AAC.37.3.377

10.1093/nar/21.8.1805

10.1128/jb.172.12.7260-7262.1990

10.1093/nar/10.1.141

10.1128/AAC.35.4.622

Studier W.F., 1990, Use of T7 RNA polymerase to direct expression of cloned genes, Meth Enzymol, 185, 89

10.1016/0022-2836(87)90479-7

10.1111/j.1365-2958.1993.tb01593.x

10.1128/jb.173.18.5854-5860.1991

Wang J.C., 1990, DNA Topology and its Biological Effects, 321

10.1128/AAC.37.3.457

10.1038/351624a0

10.1093/nar/9.24.6647

10.1128/AAC.34.6.1271

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Molecular Microbiology

Tập 13 Số 4

641-653

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