title: Comparative analysis of Histophilus somni immunoglobulin-binding protein A (IbpA) with other fic domain-containing enzymes reveals differences in substrate and nucleotide specificities.
abstract: A new family of adenylyltransferases, defined by the presence of a Fic domain, was recently discovered to catalyze the addition of adenosine monophosphate (AMP) to Rho GTPases (Yarbrough, M. L., Li, Y., Kinch, L. N., Grishin, N. V., Ball, H. L., and Orth, K. (2009) Science 323, 269-272; Worby, C. A., Mattoo, S., Kruger, R. P., Corbeil, L. B., Koller, A., Mendez, J. C., Zekarias, B., Lazar, C., and Dixon, J. E. (2009) Mol. Cell 34, 93-103). This adenylylation event inactivates Rho GTPases by preventing them from binding to their downstream effectors. We reported that the Fic domain(s) of the immunoglobulin-binding protein A (IbpA) from the pathogenic bacterium Histophilus somni adenylylates mammalian Rho GTPases, RhoA, Rac1, and Cdc42, thereby inducing host cytoskeletal collapse, which allows H. somni to breach alveolar barriers and cause septicemia. The IbpA-mediated adenylylation occurs on a functionally critical tyrosine in the switch 1 region of these GTPases. Here, we conduct a detailed characterization of the IbpA Fic2 domain and compare its activity with other known Fic adenylyltransferases, VopS (Vibrio outer protein S) from the bacterial pathogen Vibrio parahaemolyticus and the human protein HYPE (huntingtin yeast interacting protein E; also called FicD). We also included the Fic domains of the secreted protein, PfhB2, from the opportunistic pathogen Pasteurella multocida, in our analysis. PfhB2 shares a common domain architecture with IbpA and contains two Fic domains. We demonstrate that the PfhB2 Fic domains also possess adenylyltransferase activity that targets the switch 1 tyrosine of Rho GTPases. Comparative kinetic and phylogenetic analyses of IbpA-Fic2 with the Fic domains of PfhB2, VopS, and HYPE reveal important aspects of their specificities for Rho GTPases and nucleotide usage and offer mechanistic insights for determining nucleotide and substrate specificities for these enzymes. Finally, we compare the evolutionary lineages of Fic proteins with those of other known adenylyltransferases.
authors.affiliation: Department of Pharmacology, Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093-0721, USA. smattoo@ucsd.edu
authors.forename: Seema
authors.initials: S
authors.lastname: Mattoo
authors.forename: Eric
authors.initials: E
authors.lastname: Durrant
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Chen
authors.forename: Junyu
authors.initials: J
authors.lastname: Xiao
authors.forename: Cheri S
authors.initials: CS
authors.lastname: Lazar
authors.forename: Gerard
authors.initials: G
authors.lastname: Manning
authors.forename: Jack E
authors.initials: JE
authors.lastname: Dixon
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Worby
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D019143:Evolution, Molecular; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics; D015661:Pasteurellaceae / Q000201:enzymology* / Q000235:genetics; D017434:Protein Structure, Tertiary; D014736:Vibrio parahaemolyticus / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D009713:Nucleotidyltransferases