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Infection and Immunity, December 2001, p. 7326-7333, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7326-7333.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evidence that Mycobacterial PE_PGRS Proteins Are Cell Surface Constituents That Influence Interactions with Other Cells

Michael J. Brennan,1,* Giovanni Delogu,1,dagger Yiping Chen,1,Dagger Stoyan Bardarov,2 Jordan Kriakov,2 Mohammad Alavi,1,§ and William R. Jacobs Jr.2

Laboratory of Mycobacterial Diseases and Cellular Immunology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892,1 and Howard Hughes Medical Institute, Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 104612

Received 24 January 2001/Returned for modification 3 April 2001/Accepted 11 September 2001

The elucidation of the genomic sequence of Mycobacterium tuberculosis revealed the presence of a novel multigene family designated PE/PE_PGRS that encodes numerous, highly related proteins of unknown function. In this study, we demonstrate that a transposon insertion in a PE_PGRS gene (1818PE_PGRS) found in Mycobacterium bovis BCG Pasteur, which is the BCG homologue of the M. tuberculosis H37Rv gene Rv1818c, introduces new phenotypic properties to this BCG strain. These properties include dispersed growth in liquid medium and reduced infection of macrophages. Complementation of the 1818PE_PGRS::Tn5367 mutant with the wild-type gene restores both aggregative growth (clumping) in liquid medium and reestablishes infectivity of macrophages to levels equivalent to those for the parent BCG strain. Western blot analysis using antisera raised against the 1818PE_PGRS protein shows that PE_PGRS proteins are found in cell lysates of BCG and M. tuberculosis H37Ra and in the cell wall fraction of M. tuberculosis H37Rv. Moreover, immunofluorescent labeling of mycobacteria indicates that certain PE_PGRS proteins are localized at the cell surface of BCG and M. tuberculosis. Together these results suggest that certain PE_PGRS proteins may be found at the surface of mycobacteria and influence both cell surface interactions among mycobacteria as well as the interactions of mycobacteria with macrophages.

* Corresponding author. Mailing address: CBER/FDA, Building 29, Room 502, 29 Lincoln Dr. (HFM-431), Bethesda, MD 20892. Phone: (301) 496-9559. Fax: (301) 402-2776. E-mail: Brennan{at}cber.fda.gov.

dagger Present address: Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/b, 07100 Sassari, Italy.

Dagger Present address: Department of Otologic Research, College of Medicine and Public Health, The Ohio State University, Columbus, OH 43210.

§ Present address: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202.

Infection and Immunity, December 2001, p. 7326-7333, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7326-7333.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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