Localization of P42 and F 1 -ATPase α-Subunit Homolog of the Gliding Machinery in Mycoplasma mobile Revealed by Newly Developed Gene Manipulation and Fluorescent Protein Tagging
Author(s) -
Isil Tulum,
Masaru Yabe,
Atsuko Uenoyama,
Makoto Miyata
Publication year - 2014
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.01418-13
Subject(s) - biology , protein subunit , atpase , microbiology and biotechnology , transposable element , peptide sequence , gene , genetics , biochemistry , mutant , enzyme
Mycoplasma mobile has a unique mechanism that enables it to glide on solid surfaces faster than any other gliding mycoplasma. To elucidate the gliding mechanism, we developed a transformation system forM. mobile based on a transposon derived from Tn4001 . Modification of the electroporation conditions, outgrowth time, and colony formation from the standard method forMycoplasma species enabled successful transformation. A fluorescent-protein tagging technique was developed using the enhanced yellow fluorescent protein (EYFP) and applied to two proteins that have been suggested to be involved in the gliding mechanism: P42 (MMOB1050), which is transcribed as continuous mRNA with other proteins essential for gliding, and a homolog of the F1 -ATPase α-subunit (MMOB1660). Analysis of the amino acid sequence of P42 by PSI-BLAST suggested that P42 evolved from a common ancestor with FtsZ, the bacterial tubulin homologue. The roles of P42 and the F1 -ATPase subunit homolog are discussed as part of our proposed gliding mechanism.
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