Legionella eukaryotic-like type IV substrates interfere with organelle trafficking

被引:222
作者
de Felipe, Karim Suwwan [1 ,2 ]
Glover, Robert T. [2 ]
Charpentier, Xavier [2 ]
Anderson, O. Roger [3 ]
Reyes, Moraima
Pericone, Christopher D. [2 ]
Shuman, Howard A. [1 ,2 ]
机构
[1] Columbia Univ, Med Ctr, Integrated Program Cellular Mol & Biophys Studies, New York, NY 10027 USA
[2] Columbia Univ, Med Ctr, Dept Microbiol, New York, NY USA
[3] Columbia Univ, Lamont Doherty Earth Observ, Div Biol & Paleo Environm, Palisades, NY USA
关键词
D O I
10.1371/journal.ppat.1000117
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Legionella pneumophila, the causative agent of Legionnaires' disease, evades phago-lysosome fusion in mammalian and protozoan hosts to create a suitable niche for intracellular replication. To modulate vesicle trafficking pathways, L. pneumophila translocates effector proteins into eukaryotic cells through a Type IVB macro-molecular transport system called the Icm-Dot system. In this study, we employed a fluorescence-based translocation assay to show that 33 previously identified Legionella eukaryotic-like genes (leg) encode substrates of the Icm-Dot secretion system. To assess which of these proteins may contribute to the disruption of vesicle trafficking, we expressed each gene in yeast and looked for phenotypes related to vacuolar protein sorting. We found that LegC3-GFP and LegC7/YlfA-GFP caused the mis-secretion of CPY-Invertase, a fusion protein normally restricted to the yeast vacuole. We also found that LegC7/YlfA-GFP and its paralog LegC2/YlfB-GFP formed large structures around the yeast vacuole while LegC3-GFP localized to the plasma membrane and a fragmented vacuole. In mammalian cells, LegC2/YlfB-GFP and LegC7/YlfA-GFP were found within large structures that colocalized with anti-KDEL antibodies but excluded the lysosomal marker LAMP-1, similar to what is observed in Legionella-containing vacuoles. LegC3-GFP, in contrast, was observed as smaller structures which had no obvious co-localization with KDEL or LAMP-1. Finally, LegC3-GFP caused the accumulation of many endosome-like structures containing undigested material when expressed in the protozoan host Dictyostelium discoideum. Our results demonstrate that multiple Leg proteins are Icm/Dot-dependent substrates and that LegC3, LegC7/YlfA, and LegC2/YlfB may contribute to the intracellular trafficking of L. pneumophila by interfering with highly conserved pathways that modulate vesicle maturation.
引用
收藏
页数:16
相关论文
共 75 条
[51]   The Legionella IcmS-IcmW protein complex is important for Dot/Icm-mediated protein translocation [J].
Ninio, S ;
Zuckman-Cholon, DM ;
Cambronne, ED ;
Roy, CR .
MOLECULAR MICROBIOLOGY, 2005, 55 (03) :912-926
[52]   Phenotypic effects of membrane protein overexpression in Saccharomyces cerevisiae [J].
Osterberg, Marie ;
Kim, Hyun ;
Warringer, Jonas ;
Melen, Karin ;
Blomberg, Anders ;
von Heijne, Gunnar .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (30) :11148-11153
[53]   Intra-amoebal development of Legionella pneumophila and the potential role of amoebae in the transmission of Legionnaires' disease [J].
Philippe, C. ;
Blech, M. F. ;
Hartemann, P. .
MEDECINE ET MALADIES INFECTIEUSES, 2006, 36 (04) :196-200
[54]   MORPHOLOGICAL CLASSIFICATION OF THE YEAST VACUOLAR PROTEIN SORTING MUTANTS - EVIDENCE FOR A PREVACUOLAR COMPARTMENT IN CLASS-E VPS MUTANTS [J].
RAYMOND, CK ;
HOWALDSTEVENSON, I ;
VATER, CA ;
STEVENS, TH .
MOLECULAR BIOLOGY OF THE CELL, 1992, 3 (12) :1389-1402
[55]   Attachment and fusion of endoplasmic reticulum with vacuoles containing Legionella pneumophila [J].
Robinson, CG ;
Roy, CR .
CELLULAR MICROBIOLOGY, 2006, 8 (05) :793-805
[56]   PROTEIN SORTING IN SACCHAROMYCES-CEREVISIAE - ISOLATION OF MUTANTS DEFECTIVE IN THE DELIVERY AND PROCESSING OF MULTIPLE VACUOLAR HYDROLASES [J].
ROBINSON, JS ;
KLIONSKY, DJ ;
BANTA, LM ;
EMR, SD .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (11) :4936-4948
[57]   A SACCHAROMYCES-CEREVISIAE GENOMIC PLASMID BANK BASED ON A CENTROMERE-CONTAINING SHUTTLE VECTOR [J].
ROSE, MD ;
NOVICK, P ;
THOMAS, JH ;
BOTSTEIN, D ;
FINK, GR .
GENE, 1987, 60 (2-3) :237-243
[59]   Legionella pneumophila DotA protein is required for early phagosome trafficking decisions that occur within minutes of bacterial uptake [J].
Roy, CR ;
Berger, KH ;
Isberg, RR .
MOLECULAR MICROBIOLOGY, 1998, 28 (03) :663-674
[60]   IDENTIFICATION OF LEGIONELLA-PNEUMOPHILA GENES REQUIRED FOR GROWTH WITHIN AND KILLING OF HUMAN MACROPHAGES [J].
SADOSKY, AB ;
WIATER, LA ;
SHUMAN, HA .
INFECTION AND IMMUNITY, 1993, 61 (12) :5361-5373