title: Lack of the consensus sequence necessary for tryptophan prenylation in the ComX pheromone precursor.
abstract: ComX, an oligopeptide pheromone that stimulates the natural genetic competence controlled by quorum sensing in Bacillus subtilis and related bacilli, contains a prenyl-modified tryptophan residue. Since ComX is the only protein known to contain prenylated tryptophan, the universality of this unique posttranslational modification has yet to be determined. Recently, we developed a cell-free assay system in which the tryptophan residue in the ComX(RO-E-2) pheromone precursor derived from B. subtilis strain RO-E-2 can be geranylated by the ComQ(RO-E-2) enzyme. We report here our attempt to identify the consensus sequence surrounding the geranylated tryptophan residue by using the cell-free system with various ComX(RO-E-2) pheromone precursor analogs. We found that [47-58]ComX(RO-E-2), corresponding to the C-terminal 12-residue peptide of the pheromone precursor, contained a short sequence essential for geranylation. We also found that the length of the sequence between the tryptophan residue and the C-terminus was important for geranylation, and that some [47-58]ComX(RO-E-2) pheromone precursor amino acids were involved in the geranylation reaction. However, we could not identify a consensus sequence surrounding the geranylated tryptophan. Our evidence suggests that, like Rab which lacks a consensus sequence yet is geranylgeranyl-modified on a cysteine residue, the ComX pheromone and its precursor also lack a consensus sequence.
authors.affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi 464-8601, Japan. fumitada_tsuji@ajinomoto.com
authors.forename: Fumitada
authors.initials: F
authors.lastname: Tsuji
authors.forename: Ayako
authors.initials: A
authors.lastname: Ishihara
authors.forename: Aya
authors.initials: A
authors.lastname: Nakagawa
authors.forename: Masahiro
authors.initials: M
authors.lastname: Okada
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Kitamura
authors.forename: Kyoko
authors.initials: K
authors.lastname: Kanamaru
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Masuda
authors.forename: Kazuma
authors.initials: K
authors.lastname: Murakami
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Irie
authors.forename: Youji
authors.initials: Y
authors.lastname: Sakagami
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002474:Cell-Free System; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004926:Escherichia coli / Q000235:genetics; D056084:Genetic Fitness; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010675:Pheromones / Q000235:genetics* / Q000378:metabolism; D054876:Prenylation; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D053038:Quorum Sensing / Q000235:genetics; D014364:Tryptophan / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C087040:ComX protein, Bacillus subtilis; D008565:Membrane Proteins; D010675:Pheromones; D020033:Protein Isoforms; D011498:Protein Precursors; C070739:comQ protein, Bacillus subtilis; D014364:Tryptophan