Calponin h1-S175T point mutation enhances resistance to actin cytoskeleton perturbation in human cancer cells.
To investigate Calponin h1 (CNh1) regions responsible for suppressing cancer cell motility, the interaction between CNh1 and actin in HeLa cells was examined. First, it was observed that the actin binding of CNh1 depends on the calponin repeat 1 (CNR 1), region more than the actin binding site region. Next, point mutantions were generated at S175 and/or T184 in CNR1, substrates of protein kinase C, and it was observed by cellular immunostaining that the actin binding of CNh1 depends on 175th amino acid. Furthermore, the point mutation S175T exhibited more resistance to actin rearrangement by cytochalasin D and PDBu than intact CNh1, suppressing cell motility induced by PDBu. This result indicates that S175T may be an effective target for new cancer treatments.
Department of Molecular Oncology, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, Matsumoto, Japan. stangch@shinshu-u.ac.jp
Akemi
A
Sugaya
Michiko
M
Takeoka
Naoki
N
Itano
Ayako
A
Taguchi
Takashi
T
Ehara
Shun'ichiro
S
Taniguchi
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D019943:Amino Acid Substitution; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics; D015638:Cytochalasin D / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D017354:Point Mutation; D012694:Serine / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D000096985:Calponins
D000199:Actins; D002135:Calcium-Binding Proteins; D015638:Cytochalasin D; D008840:Microfilament Proteins; D015240:Phorbol 12,13-Dibutyrate; D012694:Serine; D013912:Threonine; D000096985:Calponins