Anaplasma phagocytophilum Rab10-dependent parasitism of the trans-Golgi network is critical for completion of the infection cycle.
Anaplasma phagocytophilum is an emerging human pathogen and obligate intracellular bacterium. It inhabits a host cell-derived vacuole and cycles between replicative reticulate cell (RC) and infectious dense-cored (DC) morphotypes. Host-pathogen interactions that are critical for RC-to-DC conversion are undefined. We previously reported that A. phagocytophilum recruits green fluorescent protein (GFP)-tagged Rab10, a GTPase that directs exocytic traffic from the sphingolipid-rich trans-Golgi network (TGN) to its vacuole in a guanine nucleotide-independent manner. Here, we demonstrate that endogenous Rab10-positive TGN vesicles are not only routed to but also delivered into the A. phagocytophilum-occupied vacuole (ApV). Consistent with this finding, A. phagocytophilum incorporates sphingolipids while intracellular and retains them when naturally released from host cells. TGN vesicle delivery into the ApV is Rab10 dependent, up-regulates expression of the DC-specific marker, APH1235, and is critical for the production of infectious progeny. The A. phagocytophilum surface protein, uridine monophosphate kinase, was identified as a guanine nucleotide-independent, Rab10-specific ligand. These data delineate why Rab10 is important for the A. phagocytophilum infection cycle and expand the understanding of the benefits that exploiting host cell membrane traffic affords intracellular bacterial pathogens.
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Hilary K
HK
Truchan
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Lauren
L
VieBrock
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Chelsea L
CL
Cockburn
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Nore
N
Ojogun
Molecular Biology and Genetics Program, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Brian P
BP
Griffin
Department of Surgery, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|Department of Biochemistry and Molecular Biology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Dayanjan S
DS
Wijesinghe
Department of Biochemistry and Molecular Biology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|Massey Cancer Center, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|The Victoria Johnson Center, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|Institute for Molecular Medicine, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|Research and Development, Hunter Holmes McGuire Veterans Administration Medical Center, Richmond, VA, USA.
Charles E
CE
Chalfant
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.|Molecular Biology and Genetics Program, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Jason A
JA
Carlyon
D041081:Anaplasma phagocytophilum / Q000254:growth & development*; D002460:Cell Line; D022162:Cytoplasmic Vesicles / Q000737:chemistry / Q000378:metabolism*; D006790:Host-Parasite Interactions; D006801:Humans; D009703:Nucleoside-Phosphate Kinase / Q000378:metabolism; D011485:Protein Binding; D014617:Vacuoles / Q000382:microbiology*; D020691:rab GTP-Binding Proteins / Q000032:analysis*; D021601:trans-Golgi Network / Q000382:microbiology*
C023276:uridine monophosphate kinase; D009703:Nucleoside-Phosphate Kinase; C538901:Rab10 protein, human; D020691:rab GTP-Binding Proteins
Anaplasmataceae; UMPK; intracellular parasitism; lipidomics; nutritional virulence; obligate intracellular