Analysis of type IV pilus and its associated motility in Myxococcus xanthus using an antibody reactive with native pilin and pili.
Myxococcus xanthus possesses a social gliding motility that requires type IV pili (TFP). According to the current model, M. xanthus pili attach to an external substrate and retract, pulling the cell body forward along their long axis. By analogy with the situation in other bacteria employing TFP-dependent motility, M. xanthus pili have been assumed to be composed of pilin (PilA) subunits, but this has not previously been confirmed. The first 28 amino acids of the M. xanthus PilA protein share extensive homology with the N-terminal oligomerization domain of pilins in other bacterial species. To facilitate purification, the authors engineered a truncated form of M. xanthus PilA lacking the first 28 amino acids and purified this protein in soluble form. Polyclonal antibody generated against this protein was reactive with native pilin and pili. Using this antibody, it was confirmed that TFP of M. xanthus are indeed composed of PilA, and that TFP are located unipolarly and required for social gliding motility via retraction. Using tethering as well as motility assays, details of pili function in M. xanthus social motility were further examined.
Li
Yinuo
Y
Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
Lux
Renate
R
School of Dentistry, University of California, Los Angeles, CA 90095, USA.
Pelling
Andrew E
AE
Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA.
Gimzewski
James K
JK
Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA.
Shi
Wenyuan
W
School of Dentistry, University of California, Los Angeles, CA 90095, USA.|Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D036781:Fimbriae Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009068:Movement; D016941:Myxococcus xanthus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D016415:Sequence Alignment
D000907:Antibodies, Bacterial; D036781:Fimbriae Proteins