Abstract
Tn4001 mutagenesis identified a new competence gene in Streptococcus gordonii Challis designated comYA. A comYA mutant was completely deficient in transformation and exhibited decreased levels of DNA binding and hydrolysis. The deduced 319-amino-acid ComYA protein exhibited 57% similarity and 33% identity to the ComGA transporter protein of Bacillus subtilis and contained the Walker A-box motif conserved in ATP-binding proteins as well as aspartic acid boxes Asp-1 and Asp-2 present in some components of the general secretory pathway of gram-negative bacteria. comYA appeared to be part of a putative operon encompassing a comGB homolog, designated comYB, together with sequences that could encode ComGC- and ComGD-like peptides designated ComYC and ComYD, respectively, as well as other components. The putative ComYC and ComYD peptides had leader sequences similar to the type IV N-methylphenylalanine pilins of gram-negative bacteria, but unlike other examples in this class, including B. subtilis, they contained an alanine at position -1 of the leader instead of the usual glycine residue. Northern analysis identified a single 6.0-kb comYA-containing transcript strictly dependent on exogenous competence factor for expression in ComA1 cells. An identical pattern of expression was seen in wild-type Challis cells grown under conditions of maximal competence but not in cells that were noncompetent.
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- Albano M., Breitling R., Dubnau D. A. Nucleotide sequence and genetic organization of the Bacillus subtilis comG operon. J Bacteriol. 1989 Oct;171(10):5386–5404. doi: 10.1128/jb.171.10.5386-5404.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boor K. J., Duncan M. L., Price C. W. Genetic and transcriptional organization of the region encoding the beta subunit of Bacillus subtilis RNA polymerase. J Biol Chem. 1995 Sep 1;270(35):20329–20336. doi: 10.1074/jbc.270.35.20329. [DOI] [PubMed] [Google Scholar]
- Breitling R., Dubnau D. A membrane protein with similarity to N-methylphenylalanine pilins is essential for DNA binding by competent Bacillus subtilis. J Bacteriol. 1990 Mar;172(3):1499–1508. doi: 10.1128/jb.172.3.1499-1508.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burrows L. L., Olah-Winfield E., Lo R. Y. Molecular analysis of the leukotoxin determinants from Pasteurella haemolytica serotypes 1 to 16. Infect Immun. 1993 Dec;61(12):5001–5007. doi: 10.1128/iai.61.12.5001-5007.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Byrne M. E., Rouch D. A., Skurray R. A. Nucleotide sequence analysis of IS256 from the Staphylococcus aureus gentamicin-tobramycin-kanamycin-resistance transposon Tn4001. Gene. 1989 Sep 30;81(2):361–367. doi: 10.1016/0378-1119(89)90197-2. [DOI] [PubMed] [Google Scholar]
- Cheng Q., Campbell E. A., Naughton A. M., Johnson S., Masure H. R. The com locus controls genetic transformation in Streptococcus pneumoniae. Mol Microbiol. 1997 Feb;23(4):683–692. doi: 10.1046/j.1365-2958.1997.2481617.x. [DOI] [PubMed] [Google Scholar]
- Chung Y. S., Dubnau D. ComC is required for the processing and translocation of comGC, a pilin-like competence protein of Bacillus subtilis. Mol Microbiol. 1995 Feb;15(3):543–551. doi: 10.1111/j.1365-2958.1995.tb02267.x. [DOI] [PubMed] [Google Scholar]
- Clewell D. B., Flannagan S. E., Jaworski D. D. Unconstrained bacterial promiscuity: the Tn916-Tn1545 family of conjugative transposons. Trends Microbiol. 1995 Jun;3(6):229–236. doi: 10.1016/s0966-842x(00)88930-1. [DOI] [PubMed] [Google Scholar]
- Deddish P., Slade H. D. Binding of deoxyribonucleic acid by cell walls of transformable and nontransformable streptococci. J Bacteriol. 1971 Mar;105(3):779–786. doi: 10.1128/jb.105.3.779-786.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gu J., Ren K., Dubner R., Iadarola M. J. Cloning of a DNA binding protein that is a tyrosine kinase substrate and recognizes an upstream initiator-like sequence in the promoter of the preprodynorphin gene. Brain Res Mol Brain Res. 1994 Jul;24(1-4):77–88. doi: 10.1016/0169-328x(94)90120-1. [DOI] [PubMed] [Google Scholar]
- Hobbs M., Mattick J. S. Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes. Mol Microbiol. 1993 Oct;10(2):233–243. doi: 10.1111/j.1365-2958.1993.tb01949.x. [DOI] [PubMed] [Google Scholar]
- Håvarstein L. S., Gaustad P., Nes I. F., Morrison D. A. Identification of the streptococcal competence-pheromone receptor. Mol Microbiol. 1996 Aug;21(4):863–869. doi: 10.1046/j.1365-2958.1996.521416.x. [DOI] [PubMed] [Google Scholar]
- Leonard C. G., Cole R. M. Purification and properties of Streptococcal competence factor isolated from chemically defined medium. J Bacteriol. 1972 Apr;110(1):273–280. doi: 10.1128/jb.110.1.273-280.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leonard C. G. Early events in development of streptococcal competence. J Bacteriol. 1973 Jun;114(3):1198–1205. doi: 10.1128/jb.114.3.1198-1205.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindler L. E., Macrina F. L. Characterization of genetic transformation in Streptococcus mutans by using a novel high-efficiency plasmid marker rescue system. J Bacteriol. 1986 May;166(2):658–665. doi: 10.1128/jb.166.2.658-665.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lunsford R. D. A Tn4001 delivery system for Streptococcus gordonii (Challis). Plasmid. 1995 Mar;33(2):153–157. doi: 10.1006/plas.1995.1016. [DOI] [PubMed] [Google Scholar]
- Lunsford R. D., London J. Natural genetic transformation in Streptococcus gordonii: comX imparts spontaneous competence on strain wicky. J Bacteriol. 1996 Oct;178(19):5831–5835. doi: 10.1128/jb.178.19.5831-5835.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lunsford R. D., Macrina F. L. Molecular cloning and characterization of scrB, the structural gene for the Streptococcus mutans phosphoenolpyruvate-dependent sucrose phosphotransferase system sucrose-6-phosphate hydrolase. J Bacteriol. 1986 May;166(2):426–434. doi: 10.1128/jb.166.2.426-434.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lunsford R. D., Nguyen N., London J. DNA-binding activities in Streptococcus gordonii: identification of a receptor-nickase and a histonelike protein. Curr Microbiol. 1996 Feb;32(2):95–100. doi: 10.1007/s002849900017. [DOI] [PubMed] [Google Scholar]
- Lunsford R. D. Recovery of RNA from oral streptococci. Biotechniques. 1995 Mar;18(3):412–414. [PubMed] [Google Scholar]
- Magnuson R., Solomon J., Grossman A. D. Biochemical and genetic characterization of a competence pheromone from B. subtilis. Cell. 1994 Apr 22;77(2):207–216. doi: 10.1016/0092-8674(94)90313-1. [DOI] [PubMed] [Google Scholar]
- Pestova E. V., Håvarstein L. S., Morrison D. A. Regulation of competence for genetic transformation in Streptococcus pneumoniae by an auto-induced peptide pheromone and a two-component regulatory system. Mol Microbiol. 1996 Aug;21(4):853–862. doi: 10.1046/j.1365-2958.1996.501417.x. [DOI] [PubMed] [Google Scholar]
- Possot O., Pugsley A. P. Molecular characterization of PulE, a protein required for pullulanase secretion. Mol Microbiol. 1994 Apr;12(2):287–299. doi: 10.1111/j.1365-2958.1994.tb01017.x. [DOI] [PubMed] [Google Scholar]
- Puyet A., Greenberg B., Lacks S. A. Genetic and structural characterization of endA. A membrane-bound nuclease required for transformation of Streptococcus pneumoniae. J Mol Biol. 1990 Jun 20;213(4):727–738. doi: 10.1016/S0022-2836(05)80259-1. [DOI] [PubMed] [Google Scholar]
- Raina J. L., Macrina F. L. A competence specific inducible protein promotes in vivo recombination in Streptococcus sanguis. Mol Gen Genet. 1982;185(1):21–29. doi: 10.1007/BF00333785. [DOI] [PubMed] [Google Scholar]
- Raina J. L., Ravin A. W. Switches in macromolecular synthesis during induction of competence for transformation of Streptococcus sanguis. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6062–6066. doi: 10.1073/pnas.77.10.6062. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whittaker C. J., Klier C. M., Kolenbrander P. E. Mechanisms of adhesion by oral bacteria. Annu Rev Microbiol. 1996;50:513–552. doi: 10.1146/annurev.micro.50.1.513. [DOI] [PubMed] [Google Scholar]