cis elements and trans factors are both important in strain-specific regulation of the leukotoxin gene in Actinobacillus actinomycetemcomitans

被引:48
作者
Kolodrubetz, D
Spitznagel, J
Wang, BN
Phillips, LH
Jacobs, C
Kraig, E
机构
[1] UNIV TEXAS,HLTH SCI CTR,DEPT PERIODONT,SAN ANTONIO,TX 78284
[2] UNIV TEXAS,HLTH SCI CTR,DEPT CELLULAR & STRUCT BIOL,SAN ANTONIO,TX 78284
关键词
D O I
10.1128/IAI.64.9.3451-3460.1996
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Actinobacillus actinomycetemcomitans, the etiologic agent of localized juvenile periodontitis, produces a potent leukotoxin that kills human neutrophils. The production of leukotoxin RNA can vary more than 50-fold among isolates of A. actinomycetemcomitans, and strains expressing high levels of leukotoxin RNA are most often found at sites of periodontal disease, To assess the relative contributions of transcription factors and promoter sequences in setting the disparate levels of leukotoxin RNA found, we have undertaken classical cis/trans analyses, First, the leukotoxin promoter regions from moderately leukotoxic (Y4) and minimally leukotoxic (ATCC 33384) strains of A, actinomycetemcomitans were cloned, sequenced, and compared with the previously sequenced leukotoxin promoter region of the high-producer strain JP2. The Y4 and ATCC 33384 promoter regions each contain a 528-bp segment that is absent from JP2, Interestingly, the analysis of various deletion constructs in A. actinomycetemcomitans indicated that Y4, despite the large insertion, initiates leukotoxin RNA synthesis at the same promoter as JP2 does, To perform cis/trans analyses, these three leukotoxin promoter regions were cloned into a plasmid upstream of the reporter gene beta-galactosidase, Each plasmid was transformed into JP2, Y4, and ATCC 33384, and the beta-galactosidase levels were determined, The results indicated that the sequences responsible for down-regulating leukotoxin RNA levels in Y4 relative to JP2 are found within the transcribed region of the Y4 leukotoxin operon, Importantly, in ATCC 33384, strain-specific trans factors and promoter sequence differences are equally significant in determining the lower levels of leukotoxin RNA, We hypothesize that either strain ATCC 33384 has a negative regulatory protein (which is missing or mutated in JP2/Y4) or that JP2 and Y4 carry an activator that is missing or mutated in ATCC 33384.
引用
收藏
页码:3451 / 3460
页数:10
相关论文
共 35 条
[1]  
Ausubel FM., 1994, Curr. Protoc. Mol. Biol
[2]  
BENSON S A, 1984, Biotechniques, V2, P66
[3]   PORE-FORMING BACTERIAL PROTEIN HEMOLYSINS (CYTOLYSINS) [J].
BRAUN, V ;
FOCARETA, T .
CRITICAL REVIEWS IN MICROBIOLOGY, 1991, 18 (02) :115-158
[4]  
Braun V, 1993, Trends Microbiol, V1, P211, DOI 10.1016/0966-842X(93)90134-D
[5]   REGULATION OF ACTINOBACILLUS-ACTINOMYCETEMCOMITANS LEUKOTOXIN EXPRESSION - ANALYSIS OF THE PROMOTER REGIONS OF LEUKOTOXIC AND MINIMALLY LEUKOTOXIC STRAINS [J].
BROGAN, JM ;
LALLY, ET ;
POULSEN, K ;
KILIAN, M ;
DEMUTH, DR .
INFECTION AND IMMUNITY, 1994, 62 (02) :501-508
[6]   CONTROL OF MUCOIDY IN PSEUDOMONAS-AERUGINOSA - TRANSCRIPTIONAL REGULATION OF ALGR AND IDENTIFICATION OF THE 2ND REGULATORY GENE, ALGQ [J].
DERETIC, V ;
KONYECSNI, WM .
JOURNAL OF BACTERIOLOGY, 1989, 171 (07) :3680-3688
[7]   IDENTIFICATION AND CHARACTERIZATION OF GENETIC CLUSTER GROUPS OF ACTINOBACILLUS-ACTINOMYCETEMCOMITANS ISOLATED FROM THE HUMAN ORAL CAVITY [J].
DIRIENZO, JM ;
MCKAY, TL .
JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (01) :75-81
[8]   MOLECULAR-CLONING OF THE PLASMID RP4 PRIMASE REGION IN A MULTI-HOST-RANGE TACP EXPRESSION VECTOR [J].
FURSTE, JP ;
PANSEGRAU, W ;
FRANK, R ;
BLOCKER, H ;
SCHOLZ, P ;
BAGDASARIAN, M ;
LANKA, E .
GENE, 1986, 48 (01) :119-131
[9]   CONJUGAL TRANSFER OF BROAD-HOST-RANGE INCOMPATIBILITY GROUP-P AND GROUP-Q PLASMIDS FROM ESCHERICHIA-COLI TO ACTINOBACILLUS-ACTINOMYCETEMCOMITANS [J].
GONCHAROFF, P ;
YIP, JKK ;
WANG, H ;
SCHREINER, HC ;
PAI, JA ;
FURGANG, D ;
STEVENS, RH ;
FIGURSKI, DH ;
FINE, DH .
INFECTION AND IMMUNITY, 1993, 61 (08) :3544-3547
[10]   MUTATIONAL ANALYSIS OF THE PUTATIVE LEUKOTOXIN TRANSPORT GENES IN ACTINOBACILLUS-ACTINOMYCETEMCOMITANS [J].
GUTHMILLER, JM ;
KOLODRUBETZ, D ;
KRAIG, E .
MICROBIAL PATHOGENESIS, 1995, 18 (05) :307-321