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Rv0216, a conserved hypothetical protein from Mycobacterium tuberculosis that is essential for bacterial survival during infection, has a double hotdog fold

Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, SE-751 24 Uppsala, Sweden

(RECEIVED March 4, 2005; FINAL REVISION April 22, 2005; ACCEPTED April 22, 2005)

The Mycobacterium tuberculosis genome contains about 4000 genes, of which approximately a third code for proteins of unknown function or are classified as conserved hypothetical proteins. We have determined the three-dimensional structure of one of these, the rv0216 gene product, which has been shown to be essential for M. tuberculosis growth in vivo. The structure exhibits the greatest similarity to bacterial and eukaryotic hydratases that catalyse the R-specific hydration of 2-enoyl coenzyme A. However, only part of the catalytic machinery is conserved in Rv0216 and it showed no activity for the substrate crotonyl-CoA. The structure of Rv0216 allows us to assign new functional annotations to a family of seven other M. tuberculosis proteins, a number if which are essential for bacterial survival during infection and growth.

Keywords: Mycobacterium tuberculosis; Rv0216; hotdog fold; protein structure; crystallography

Abbreviations: ACP, acyl carrier protein • CoA, coenzyme A • COG, cluster of orthologous groups • DHD, double hotdog • FAS, fatty acid synthase • PDB, Protein Data Bank • PMSF, phenylmethane sulfonyl fluoride • RMSD, root-mean-square distance • SeMet, seleno-methionine • SHD, single hotdog • TB, tuberculosis.

Article and publication are at http://www.proteinscience.org/cgi/doi/10.1110/ps.051442305.

Reprint requests to: T. Alwyn Jones or Kristina Bäckbro, Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Box 596, SE-751 24 Uppsala, Sweden; e-mail: alwyn{at}xray.bmc.uu.se or nina{at}xray.bmc.uu.se; fax: +46-18-536971.

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