Identification and characterization of a Streptococcus pyogenes ABC transporter with multiple specificity for metal cations

被引:81
作者
Janulczyk, R
Pallon, J
Björck, L
机构
[1] Univ Lund, Dept Cell & Mol Biol, Sect Mol Pathogenesis, S-22100 Lund, Sweden
[2] Univ Lund, Dept Nucl Phys, S-22100 Lund, Sweden
关键词
D O I
10.1046/j.1365-2958.1999.01626.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Metal ions are crucial trace elements for bacteria infecting the human host. The Lral (lipoprotein receptor-associated antigen I) transporter in Streptococcus spp. belongs to the superfamily of ABC transporters. The transporter consists of a lipoprotein, an ATP-binding protein and a hydrophobic integral membrane protein. Here, we describe a new member of the Lral family in the important human pathogen Streptococcus pyogenes. The system was identified in silico by analysis of the S. pyogenes Genome Sequencing Project. The S, pyogenes operon exhibits an atypical organization compared with equivalents in other Streptococcus spp. The presence and atypical organization of the operon was verified in a number of S, pyogenes strains of different serotypes. Transcriptional analysis of the Lral operon demonstrates a polycistronic transcription attenuated by a stable stem-loop structure, which allows the lipoprotein to be expressed in larger quantities than the other two components. The localization of the native lipoprotein at the bacterial surface was shown by proteolytic digestion of S. pyogenes bacteria and NH2-terminal sequencing of a released lipoprotein fragment. Recombinant lipoprotein was expressed as a GST fusion protein, and studies of molecular interactions with metal radioisotopes demonstrated that the protein has affinity for Zn(II), Fe(III) and Cu(II). Zn(II) and Cu(II) were found to compete for the same binding site, whereas Fe(III) uses a second site. Also, proton-induced X-ray analysis of lipoprotein samples identified iron, copper and zinc. Finally, a mutant strain lacking a functional mtsABC operon was generated and showed reduced uptake of Fe-55 and Zn-65 compared with the wild-type strain. The operon encoding this novel ABC transporter with multiple specificity for metal cations is designated mtsABC, for metal transporter of Streptococcus.
引用
收藏
页码:596 / 606
页数:11
相关论文
共 53 条
[1]   CLONING OF THE STREPTOCOCCUS-GORDONII PK488 GENE, ENCODING AN ADHESIN WHICH MEDIATES COAGGREGATION WITH ACTINOMYCES-NAESLUNDII PK606 [J].
ANDERSEN, RN ;
GANESHKUMAR, N ;
KOLENBRANDER, PE .
INFECTION AND IMMUNITY, 1993, 61 (03) :981-987
[2]   MOLECULAR-IDENTIFICATION OF AN ABC TRANSPORTER COMPLEX FOR MANGANESE - ANALYSIS OF A CYANOBACTERIAL MUTANT STRAIN IMPAIRED IN THE PHOTOSYNTHETIC OXYGEN EVOLUTION PROCESS [J].
BARTSEVICH, VV ;
PAKRASI, HB .
EMBO JOURNAL, 1995, 14 (09) :1845-1853
[3]   Zinc(II) tolerance in Escherichia coli K-12: evidence that the zntA gene (o732) encodes a cation transport ATPase [J].
Beard, SJ ;
Hashim, R ;
MembrilloHernandez, J ;
Hughes, MN ;
Poole, RK .
MOLECULAR MICROBIOLOGY, 1997, 25 (05) :883-891
[4]   The Yfe system of Yersinia pestis transports iron and manganese and is required for full virulence of plague [J].
Bearden, SW ;
Perry, RD .
MOLECULAR MICROBIOLOGY, 1999, 32 (02) :403-414
[5]   An ABC transporter system of Yersinia pestis allows utilization of chelated iron by Escherichia coli SAB11 [J].
Bearden, SW ;
Staggs, TM ;
Perry, RD .
JOURNAL OF BACTERIOLOGY, 1998, 180 (05) :1135-1147
[6]   STREPTOCOCCAL CYSTEINE PROTEINASE RELEASES BIOLOGICALLY-ACTIVE FRAGMENTS OF STREPTOCOCCAL SURFACE-PROTEINS [J].
BERGE, A ;
BJORCK, L .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (17) :9862-9867
[7]   Sequence heterogeneity of PsaA, a 37-kilodalton putative adhesin essential for virulence of Streptococcus pneumoniae [J].
Berry, AM ;
Paton, JC .
INFECTION AND IMMUNITY, 1996, 64 (12) :5255-5262
[8]   FIMA, A MAJOR VIRULENCE FACTOR ASSOCIATED WITH STREPTOCOCCUS-PARASANGUIS ENDOCARDITIS [J].
BURNETTECURLEY, D ;
WELLS, V ;
VISCOUNT, H ;
MUNRO, CL ;
FENNO, JC ;
FIVESTAYLOR, P ;
MACRINA, FL .
INFECTION AND IMMUNITY, 1995, 63 (12) :4669-4674
[10]   A METHOD TO ISOLATE RNA FROM GRAM-POSITIVE BACTERIA AND MYCOBACTERIA [J].
CHEUNG, AL ;
EBERHARDT, KJ ;
FISCHETTI, VA .
ANALYTICAL BIOCHEMISTRY, 1994, 222 (02) :511-514