Helicobacter pylori CagA causes mitotic impairment and induces chromosomal instability.
Infection with cagA-positive Helicobacter pylori is the strongest risk factor for the development of gastric carcinoma. The cagA gene product CagA, which is delivered into gastric epithelial cells, specifically binds to and aberrantly activates SHP-2 oncoprotein. CagA also interacts with and inhibits partitioning-defective 1 (PAR1)/MARK kinase, which phosphorylates microtubule-associated proteins to destabilize microtubules and thereby causes epithelial polarity defects. In light of the notion that microtubules are not only required for polarity regulation but also essential for the formation of mitotic spindles, we hypothesized that CagA-mediated PAR1 inhibition also influences mitosis. Here, we investigated the effect of CagA on the progression of mitosis. In the presence of CagA, cells displayed a delay in the transition from prophase to metaphase. Furthermore, a fraction of the CagA-expressing cells showed spindle misorientation at the onset of anaphase, followed by chromosomal segregation with abnormal division axis. The effect of CagA on mitosis was abolished by elevated PAR1 expression. Conversely, inhibition of PAR1 kinase elicited mitotic delay similar to that induced by CagA. Thus, CagA-mediated inhibition of PAR1, which perturbs microtubule stability and thereby causes microtubule-based spindle dysfunction, is involved in the prophase/metaphase delay and subsequent spindle misorientation. Consequently, chronic exposure of cells to CagA induces chromosomal instability. Our findings reveal a bifunctional role of CagA as an oncoprotein: CagA elicits uncontrolled cell proliferation by aberrantly activating SHP-2 and at the same time induces chromosomal instability by perturbing the microtubule-based mitotic spindle. The dual function of CagA may cooperatively contribute to the progression of multistep gastric carcinogenesis.
Umeda
Mayumi
M
Division of Molecular Oncology, Institute for Genetic Medicine, Sapporo 060-0815; Division of Chemistry, Graduate School of Science, Sapporo 060-0815.
Murata-Kamiya
Naoko
N
Division of Molecular Oncology, Institute for Genetic Medicine, Sapporo 060-0815; Division of Chemistry, Graduate School of Science, Sapporo 060-0815.
Saito
Yasuhiro
Y
Division of Molecular Oncology, Institute for Genetic Medicine, Sapporo 060-0815.
Ohba
Yusuke
Y
Laboratory of Pathophysiology and Signal Transduction, Graduate School of Medicine, Hokkaido University, Sapporo 060-0815.
Takahashi
Masayuki
M
Division of Chemistry, Graduate School of Science, Sapporo 060-0815.
Hatakeyama
Masanori
M
Division of Molecular Oncology, Institute for Genetic Medicine, Sapporo 060-0815; Division of Chemistry, Graduate School of Science, Sapporo 060-0815; Department of Microbiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
D000705:Anaphase; D000942:Antigens, Bacterial / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D043171:Chromosomal Instability; D005822:Genetic Vectors; D016480:Helicobacter pylori / Q000378:metabolism*; D006801:Humans; D008677:Metaphase; D018715:Microscopy, Video; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D008938:Mitosis; D008954:Models, Biological; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D014162:Transfection
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C080866:cagA protein, Helicobacter pylori; C111833:MARK3 protein, human; D017346:Protein Serine-Threonine Kinases