title: HMW1 is required for cytadhesin P1 trafficking to the attachment organelle in Mycoplasma pneumoniae.
abstract: Mycoplasma pneumoniae proteins HMW1-HMW3 collectively are essential for cytadherence, but the function or requirement for each has not been defined. Cytadherence mutant M6 lacks HMW1 because of a frameshift in hmw1 and produces a truncated adherence-associated protein P30 because of a deletion at the 3' end of p30. Genetic manipulation of this mutant was used to evaluate the role of HMW1 in cytadherence. Mutant M6 was transformed with a recombinant transposon containing a wild-type p30 allele. Transformants synthesized both truncated and full-length P30, from the resident and recombinant alleles, respectively. However, these transformants remained hemadsorption negative, suggesting that HMW1 is required for cytadherence. Wild-type M. pneumoniae cells are generally elongated, tapering to form the attachment organelle at one end of the cell. The cytadhesin protein P1 is normally densely clustered on the mycoplasma surface at this differentiated terminal structure. However, both mutant M6 and M6 transformed with recombinant p30 had a striking ovoid morphology with no tapering at the tip structure, making the attachment organelle indistinguishable. Furthermore, protein P1 was randomly distributed on the mycoplasma surface rather than clustered at a polar location. In contrast, mutant M6 transformed with a recombinant transposon expressing the wild-type hmw1 allele exhibited a near-normal morphology and localized P1 to the attachment organelle. Significantly, M6 transformed with an hmw1 gene truncated slightly at the 3' end failed to restore proper morphology or P1 localization to the attachment organelle, suggesting a functional importance to the C-terminal domain of HMW1.
authors.affiliation: Department of Microbiology, University of Georgia, Athens 30602, USA.
authors.forename: T W
authors.initials: TW
authors.lastname: Hahn
authors.forename: M J
authors.initials: MJ
authors.lastname: Willby
authors.forename: D C
authors.initials: DC
authors.lastname: Krause
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D004251:DNA Transposable Elements; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D009154:Mutation; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D014169:Transformation, Bacterial
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C099979:HMW1 protein, Mycoplasma; D008565:Membrane Proteins; C035471:adhesin, Mycoplasma pneumoniae