The Rickettsia conorii autotransporter protein Sca1 promotes adherence to nonphagocytic mammalian cells.
The pathogenesis of spotted fever group (SFG) Rickettsia species, including R. conorii and R. rickettsii, is acutely dependent on adherence to and invasion of host cells, including cells of the mammalian endothelial system. Bioinformatic analyses of several rickettsia genomes revealed the presence of a cohort of genes designated sca genes that are predicted to encode proteins with homology to autotransporter proteins of Gram-negative bacteria. Previous work demonstrated that three members of this family, rOmpA (Sca0), Sca2, and rOmpB (Sca5) are involved in the interaction with mammalian cells; however, very little was known about the function of other conserved rickettsial Sca proteins. Here we demonstrate that sca1, a gene present in nearly all SFG rickettsia genomes, is actively transcribed and expressed in R. conorii cells. Alignment of Sca1 sequences from geographically diverse SFG Rickettsia species showed that there are high degrees of sequence identity and conservation of these sequences, suggesting that Sca1 may have a conserved function. Using a heterologous expression system, we demonstrated that production of R. conorii Sca1 in the Escherichia coli outer membrane is sufficient to mediate attachment to but not invasion of a panel of cultured mammalian epithelial and endothelial cells. Furthermore, preincubation of a recombinant Sca1 peptide with host cells blocked R. conorii cell association. Together, these results demonstrate that attachment to mammalian cells can be uncoupled from the entry process and that Sca1 is involved in the adherence of R. conorii to host cells.
Riley
Sean P
SP
Department of Microbiology, University of Chicago, 920 E. 58th Street, Cummings Life Sciences Center, 707A, Chicago, IL 60637, USA.
Goh
Kenneth C
KC
Hermanas
Timothy M
TM
Cardwell
Marissa M
MM
Chan
Yvonne G Y
YG
Martinez
Juan J
JJ
D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D042783:Endothelial Cells / Q000382:microbiology; D004847:Epithelial Cells / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics / Q000472:pathogenicity; D020869:Gene Expression Profiling; D006367:HeLa Cells; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D020114:Rickettsia conorii / Q000235:genetics / Q000472:pathogenicity*; D017386:Sequence Homology, Amino Acid; D014709:Vero Cells
D018829:Adhesins, Bacterial; D026901:Membrane Transport Proteins