POSTERS - BLAST X - University of Utah

BLAST X Poster #68
SYSTEMATIC DETECTION OF PROTEINS THAT LOCALIZE AT THE ATTACHMENT
ORGANELLE REGIONS OF MYCOPLASMA PNEUMONIAE BY FLUORESCENT-PROTEIN
TAGGING
Tsuyoshi Kenri 1 , Atsuko Horino 1 , Mayumi Kubota 1 , Yuko Sasaki 1 , Daisuke Nakane 2 and
Makoto Miyata 2
1. Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious
Disease, 4-7-1, Gakuen, Musashimurayama, Tokyo, 208-0011, Japan 2. Graduate School of
Science, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585, Japan
Mycoplasma pneumoniae is a cell wall-less bacterium with minimum range of genome
size required for self-replication. It is also known as a general cause of bronchitis and
pneumonia in humans. Pathogenicity of this bacterium is largely depends on its ability to attach
to respiratory epithelial cells (cytadherence). The M. pneumoniae cells attached to cell surface
also exhibit gliding motility. The cytadherence and gliding motility are mediated by a unique cell
terminal structure of this bacterium, the attachment organelle, which is a membrane protrusion
supported by internal cytoskeletal structures. Since the attachment organelle duplicates before
cell division, the organelle formation is thought to be coordinated with cell cycles. A number of
protein components of the attachment organelle (cytadherence-related proteins) have been
identified so far (HMW1, HMW2, HMW3, P1, P30, P65, P200, P24, P41, P90(B) and P40(C)).
However, configuration and fine structure of these proteins in the organelle are not fully
understood. In addition, the studies of Triton X-100 insoluble fraction of M. pneumoniae cells,
cross-linking analysis of P1 adhesin protein and isolation of various gliding mutants are
suggesting a possibility that there are more protein factors that associate with the attachment
organelle components. In this study, to explore whether there are more proteins that localize at
the organelle, we perform systematic fluorescent-protein tagging analysis for M. pneumoniae
ORFs. At present, we have finished the localization analysis of 620 ORFs (about 90% of total
ORFs). In this analysis, we observed that about 50 ORF products including previously known 9
cytadherence-related proteins localized at the attachment organelle region. The other ORF
products identified are 10 proteins involved in DNA, RNA and protein synthesis, 20 homologs of
variety of enzymes, 2 chaperons and 15 proteins of unknown function. Probably, some of these
proteins may be the components of the attachment organelle and have function in formation of
the organelle, cytadherence, gliding motility and coordination of cell cycle.
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