Agnès Dintilhac1, Geneviève Alloing2, Yves Quentin2, Leiv Sigve Håvarstein2
1Laboratoire de Microbiologie et Génétique Moléculaire CNRS-UPR 9007, Université Paul Sabatier, Toulouse, France.
2Laboratoire de Microbiologie et Génétique moléculaire CNRS-UPR 9007, Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex (France)
Tóm tắt
The adcCBA putative operon of Streptococcus pneumoniae, an important human pathogen, was identified in a search for transformation‐deficient mutants. It was found to exhibit homology to ATP‐binding cassette (ABC) transport operons encoding streptococcal adhesins such as FimA of Streptococcus parasanguis and PsaA of S. pneumoniae. The latter was recently shown to be essential for virulence as judged by intranasal or intraperitoneal challenge of mice. We suggested previously that AdcA, together with a set of 14 proteins, including PsaA and homologous adhesins, defines a new family of external solute‐binding proteins specific for metals. In this work, Northern analysis revealed the existence of two adcB–adcA specific transcripts originating within adcC or further upstream, consistent with the hypothesis that adc is an operon. Investigation of growth of adc and psaA mutants in synthetic medium revealed that the addition of Zn improved the growth rate of the former, whereas the latter exhibited an absolute requirement for added Mn. A psaA–adc double mutant turned out to be essentially non‐viable unless both metals were added in the appropriate ratio. Taken together, these results suggest a previously undocumented requirement of S. pneumoniae for Zn and Mn. The addition of Zn also restored near‐normal spontaneous transformation of adc mutant cells in standard transformation medium. Zn was found to be specifically required soon after contact of cells with the competence‐stimulating peptide, revealing an unsuspected need for Zn in transformation of S. pneumoniae. The removal of Mn from standard transformation medium also resulted in transformation deficiency of psaA mutant cells. Taken together, these results lead us to propose that Adc is an ABC‐type Zn permease, the first such protein complex identified in any organism, and that Psa is an ABC‐type Mn permease complex.
