Host alpha-adducin is redistributed and localized to the inclusion membrane in chlamydia- and chlamydophila-infected cells.
A large-scale analysis of proteins involved in host-cell signalling pathways was performed using chlamydia-infected murine cells in order to identify host proteins that are differentially activated or localized following infection. Two proteins whose distribution was altered in Chlamydia trachomatis-infected cells relative to mock-infected cells were the actin-binding protein adducin and the regulatory kinase Raf-1. Immunoblot analysis with antibodies to both phosphorylated and non-phosphorylated forms of these proteins demonstrated that the abundance of each protein was markedly reduced in the cytosolic fraction of C. trachomatis- and Chlamydophila caviae-infected cells, but the total cellular protein abundance remained unaffected by infection. Fluorescence microscopy of chlamydia-infected cells using anti-alpha-adducin antibodies demonstrated labelling at or near the chlamydial inclusion membrane. Treatment of infected cells with nocodazole or cytochalasin D did not affect alpha-adducin that was localized to the margins of the inclusion. The demonstration of alpha-adducin and Raf-1 redistribution within cells infected by different chlamydiae provides novel opportunities for analysis of host-pathogen interactions in this system.
Chu
Hencelyn G
HG
Department of Microbiology, Oregon State University, Corvallis, OR 97331-3804, USA.|Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331-4804, USA.
Weeks
Sara K
SK
Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331-4804, USA.
Gilligan
Diana M
DM
Puget Sound Blood Center, University of Washington, Seattle, WA 98104, USA.
Rockey
Daniel D
DD
Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331-4804, USA.
D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D002692:Chlamydia trachomatis / Q000472:pathogenicity / Q000502:physiology*; D023501:Chlamydophila / Q000145:classification / Q000472:pathogenicity* / Q000502:physiology; D016993:Chlamydophila pneumoniae / Q000472:pathogenicity / Q000502:physiology; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D015151:Immunoblotting; D002479:Inclusion Bodies / Q000378:metabolism; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism*; D000099291:Adducins
D002148:Calmodulin-Binding Proteins; D019908:Proto-Oncogene Proteins c-raf; D000099291:Adducins