Molecular characterization of the gyrB region of the oral spirochete, Treponema denticola

被引：5

作者：

Greene, SR ^{[1
]}

Stamm, LV ^{[1
]}

机构：

[1] Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, Program Infect Dis, Chapel Hill, NC 27599 USA

来源：

GENE | 2000年 / 253卷 / 02期

关键词：

coumermycin A(1) resistance; DnaA boxes; DNA gyrase;

D O I：

10.1016/S0378-1119(00)00254-7

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The nucleotide (nt) sequence of the Treponema denticola (Td) DNA gyrase beta-subunit gene (gyrB) has been determined. Southern blot analysis of Td chromosomal DNA indicated that gyrB is present as a single copy. Approximately 3.2 kb of the nt sequence 5' and 0.7 kb of nucleotide sequence 3' of gyrB were obtained. Analysis of the deduced amino acid (aa) sequence revealed two complete open reading frames (ORFs) (ORF1 and ORF3) and a truncated ORF (ORF4'). ORF1 has no homology to sequences in the databases, whereas ORF3 and ORF4' have significant homology to several bacterial DnaA (replication initiator) and DnaE (DNA polymerase III) proteins respectively. RT-PCR data showed that orf1-gyrB are co-transcribed, while dnaA-dnaE are co-transcribed but in the opposite direction. These data indicated that the gene organization of the Td gyrB region is unique compared with that of other bacteria. Eighteen putative DnaA boxes with several AT-rich regions were identified in the dnaA-dnaE intergenic region, and three putative DnaA boxes were identified in the gyrB-dnaA intergenic region. Spontaneous coumermycin A(1)-resistant Td mutants were isolated and characterized. The mutants have a >20-fold higher resistance to coumermycin A(1) than wild-type Td. A single point mutation in gyrB that changed GyrB Lys(136) to Glu or Thr appears to be responsible for the coumermycin A(1) resistance. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：259 / 269

页数：11

共 36 条

[1] THE 43-KILODALTON N-TERMINAL FRAGMENT OF THE DNA GYRASE-B PROTEIN HYDROLYZES ATP AND BINDS COUMARIN DRUGS [J].

ALI, JA ;

JACKSON, AP ;

HOWELLS, AJ ;

MAXWELL, A .

BIOCHEMISTRY, 1993, 32 (10) :2717-2724

[2] Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].

Altschul, SF ;

Madden, TL ;

Schaffer, AA ;

Zhang, JH ;

Zhang, Z ;

Miller, W ;

Lipman, DJ .

NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402

[3] Mutations in Bartonella bacilliformis gyrB confer resistance to coumermycin A1 [J].

Battisti, JM ;

Smitherman, LS ;

Samuels, DS ;

Minnick, MF .

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1998, 42 (11) :2906-2913

[4] CLONING, SEQUENCING, AND EXPRESSION OF THE DNA GYRASE GENES FROM STAPHYLOCOCCUS-AUREUS [J].

BROCKBANK, SMV ;

BARTH, PT .

JOURNAL OF BACTERIOLOGY, 1993, 175 (11) :3269-3277

[5]

CASTILLO ID, 1991, P NATL ACAD SCI USA, V88, P8860

[6]

CONTRERAS A, 1992, MOL MICROBIOL, V6, P1617

[7] Genomic sequence of a Lyme disease spirochaete, Borrelia burgdorferi [J].

Fraser, CM ;

Casjens, S ;

Huang, WM ;

Sutton, GG ;

Clayton, R ;

Lathigra, R ;

White, O ;

Ketchum, KA ;

Dodson, R ;

Hickey, EK ;

Gwinn, M ;

Dougherty, B ;

Tomb, JF ;

Fleischmann, RD ;

Richardson, D ;

Peterson, J ;

Kerlavage, AR ;

Quackenbush, J ;

Salzberg, S ;

Hanson, M ;

vanVugt, R ;

Palmer, N ;

Adams, MD ;

Gocayne, J ;

Weidman, J ;

Utterback, T ;

Watthey, L ;

McDonald, L ;

Artiach, P ;

Bowman, C ;

Garland, S ;

Fujii, C ;

Cotton, MD ;

Horst, K ;

Roberts, K ;

Hatch, B ;

Smith, HO ;

Venter, JC .

NATURE, 1997, 390 (6660) :580-586

[8] Complete genome sequence of Treponema pallidum, the syphilis spirochete [J].

Fraser, CM ;

Norris, SJ ;

Weinstock, CM ;

White, O ;

Sutton, GG ;

Dodson, R ;

Gwinn, M ;

Hickey, EK ;

Clayton, R ;

Ketchum, KA ;

Sodergren, E ;

Hardham, JM ;

McLeod, MP ;

Salzberg, S ;

Peterson, J ;

Khalak, H ;

Richardson, D ;

Howell, JK ;

Chidambaram, M ;

Utterback, T ;

McDonald, L ;

Artiach, P ;

Bowman, C ;

Cotton, MD ;

Fujii, C ;

Garland, S ;

Hatch, B ;

Horst, K ;

Roberts, K ;

Sandusky, M ;

Weidman, J ;

Smith, HO ;

Venter, JC .

SCIENCE, 1998, 281 (5375) :375-388

[9] STRUCTURE OF THE DNAA REGION OF PSEUDOMONAS-PUTIDA - CONSERVATION AMONG 3 BACTERIA, BACILLUS-SUBTILIS, ESCHERICHIA-COLI AND PSEUDOMONAS-PUTIDA [J].

FUJITA, MQ ;

YOSHIKAWA, H ;

OGASAWARA, N .

MOLECULAR & GENERAL GENETICS, 1989, 215 (03) :381-387

[10] CHARACTERIZATION, CLONING, AND BINDING-PROPERTIES OF THE MAJOR 53-KILODALTON TREPONEMA-DENTICOLA SURFACE-ANTIGEN [J].

HAAPASALO, M ;

MULLER, KH ;

UITTO, VJ ;

LEUNG, WK ;

MCBRIDE, BC .

INFECTION AND IMMUNITY, 1992, 60 (05) :2058-2065

← 1 2 3 4 →