A unique nine-gene comY operon in Streptococcus mutans

Justin Merritt, Fengxia Qi, Wenyuan Shi

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Many Gram-positive and Gram-negative bacteria possess natural competence mechanisms for DNA capture and internalization. In Bacillus subtilis, natural competence is absolutely dependent upon the presence of a seven-gene operon known as the comG operon (comGA-G). In species of Streptococcus, this function has been described for a four-gene operon (comYA-D in Streptococcus gordonii and cglA-D in Streptococcus pneumoniae). In this study, a nine-orf operon (named comYA-I) required for natural competence in Streptococcus mutans was identified and characterized. Orf analysis of this operon indicates that the first four Orfs (ComYA-D) share strong homology with ComYA-D of S. gordonii and CglA-D of S. pneumoniae, the fifth to seventh Orfs (ComYE-G match conserved hypothetical proteins from various species of Streptococcus with ComYF possessing a predicted ComGF domain, the eighth Orf (ComYH) shows a strong homology to numerous DNA methyltransferases from restriction/modification systems, and the ninth Orf (ComYI) is homologous to acetate kinase (AckA). RT-PCR analysis of the orf junctions confirmed that all nine orfs were present in a single transcript, while real-time RT-PCR analysis demonstrated that these orfs were expressed at a level very similar to that of the first orf in the operon. Mutations were constructed in all nine putative orfs. The first seven genes (comYA-G) were found to be essential for natural competence, while comYH and comYI had reduced and normal natural competence ability, respectively. Analyses of S. mutans comY-luciferase reporter fusions indicated that comY expression is growth-phase dependent, with maximal expression at an OD600 of about 0.2, while mutations in ciaH, comC and luxS reduced the level of comY expression. In addition, comY operon expression appears to be correlated with natural competence ability.

Original languageEnglish (US)
Pages (from-to)157-166
Number of pages10
JournalMicrobiology
Volume151
Issue number1
DOIs
StatePublished - Jan 2005
Externally publishedYes

Fingerprint

Contagious Ecthyma
Streptococcus mutans
Operon
Mental Competency
Genes
Streptococcus gordonii
Aptitude
Streptococcus pneumoniae
Streptococcus
Acetate Kinase
DNA Restriction-Modification Enzymes
Mutation
DNA
Methyltransferases
Bacillus subtilis
Gram-Negative Bacteria
Luciferases
Real-Time Polymerase Chain Reaction

ASJC Scopus subject areas

  • Microbiology

Cite this

A unique nine-gene comY operon in Streptococcus mutans. / Merritt, Justin; Qi, Fengxia; Shi, Wenyuan.

In: Microbiology, Vol. 151, No. 1, 01.2005, p. 157-166.

Research output: Contribution to journalArticle

Merritt, Justin ; Qi, Fengxia ; Shi, Wenyuan. / A unique nine-gene comY operon in Streptococcus mutans. In: Microbiology. 2005 ; Vol. 151, No. 1. pp. 157-166.
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abstract = "Many Gram-positive and Gram-negative bacteria possess natural competence mechanisms for DNA capture and internalization. In Bacillus subtilis, natural competence is absolutely dependent upon the presence of a seven-gene operon known as the comG operon (comGA-G). In species of Streptococcus, this function has been described for a four-gene operon (comYA-D in Streptococcus gordonii and cglA-D in Streptococcus pneumoniae). In this study, a nine-orf operon (named comYA-I) required for natural competence in Streptococcus mutans was identified and characterized. Orf analysis of this operon indicates that the first four Orfs (ComYA-D) share strong homology with ComYA-D of S. gordonii and CglA-D of S. pneumoniae, the fifth to seventh Orfs (ComYE-G match conserved hypothetical proteins from various species of Streptococcus with ComYF possessing a predicted ComGF domain, the eighth Orf (ComYH) shows a strong homology to numerous DNA methyltransferases from restriction/modification systems, and the ninth Orf (ComYI) is homologous to acetate kinase (AckA). RT-PCR analysis of the orf junctions confirmed that all nine orfs were present in a single transcript, while real-time RT-PCR analysis demonstrated that these orfs were expressed at a level very similar to that of the first orf in the operon. Mutations were constructed in all nine putative orfs. The first seven genes (comYA-G) were found to be essential for natural competence, while comYH and comYI had reduced and normal natural competence ability, respectively. Analyses of S. mutans comY-luciferase reporter fusions indicated that comY expression is growth-phase dependent, with maximal expression at an OD600 of about 0.2, while mutations in ciaH, comC and luxS reduced the level of comY expression. In addition, comY operon expression appears to be correlated with natural competence ability.",
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