TY - JOUR
T1 - PACSIN 2 represses cellular migration through direct association with cyclin D1 but not its alternate splice form cyclin D1b
AU - Meng, Hui
AU - Tian, Lifeng
AU - Zhou, Jie
AU - Li, Zhiping
AU - Jiao, Xuanmao
AU - Li, Wayne W.
AU - Plomann, Markus
AU - Xu, Zhishun
AU - Lisanti, Michael P.
AU - Wang, Chenguang
AU - Pestell, Richard G.
N1 - Funding Information:
Protein identification by MS/MS. Protein complexes were This work was supported in part by awards from the Susan purified from whole-cell extracts prepared in lysis buffer (50 Komen Breast Cancer Foundation [BCTR0504227 to C.W.], mM Tris HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Triton National Institutes of Health [R01CA70896, R01CA75503 X-100) supplemented with protease inhibitor on affinity column and R01CA86072 to R.G.P.]. Work conducted at the Kimmel packed with M2-agarose (Sigma). Following 10–20 column Cancer Center was supported by the NIH Cancer Center Core volumes of TBS (50 mM Tris HCl, pH 7.4, 150 mM NaCl) grant [P30CA56036 to R.G.P.]. This project was partially sup-washes, protein complexes were eluted with TBS buffer con-ported by the China Scholarship Council. This project is funded taining 100 μg/mL 3x FLAG peptide. Purified complexes were in part by the Pennsylvania Department of Health grant [C.W., concentrated with Microcon YM-10 (Bedford, MA), resolved R.G.P.]. The Department specifically disclaims responsibility for on 10% SDS-PAGE gel prior to preparation for MALDI mass any analyses, interpretations or conclusions.
PY - 2011/1/1
Y1 - 2011/1/1
N2 - Cyclin D1 overexpression is a common feature of many human malignancies. Genomic deletion analysis has demonstrated a key role for cyclin D1 in cellular proliferation, angiogenesis and cellular migration. To investigate the mechanisms contributing to cyclin D1 functions, we purified cyclin D1a-associated complexes by affinity chromatography and identified the PACSIN 2 (protein kinase C and casein kinase substrate in neurons 2) protein by mass spectrometry. The PACSIN 2, but not the related PACSIN 1 and 3, directly bound wild-type cyclin D1 (cyclin D1a) at the carboxyl terminus and failed to bind cyclin D1b, the alternative splicing variant of cyclin D1. PACSIN 2 knockdown induced cellular migration and reduced cell spreading in LNCaP cells expressing cyclin D1a. In cyclin D1-/- mouse embryonic fibroblasts (MeFs), cyclin D1a, but not cyclin D1b, reduced the cell spreading to a polarized morphology. sipACSIN 2 had no effect on cellular migration of cyclin D1 -/- MeFs. Cyclin D1a restored the migratory ability of cyclin D1 -/- MeFs, which was further enhanced by knocking down PACSIN 2 with siRNA. the cyclin D1-associated protein, PACSIN 2, regulates cell spreading and migration, which are dependent on cyclin D1 expression.
AB - Cyclin D1 overexpression is a common feature of many human malignancies. Genomic deletion analysis has demonstrated a key role for cyclin D1 in cellular proliferation, angiogenesis and cellular migration. To investigate the mechanisms contributing to cyclin D1 functions, we purified cyclin D1a-associated complexes by affinity chromatography and identified the PACSIN 2 (protein kinase C and casein kinase substrate in neurons 2) protein by mass spectrometry. The PACSIN 2, but not the related PACSIN 1 and 3, directly bound wild-type cyclin D1 (cyclin D1a) at the carboxyl terminus and failed to bind cyclin D1b, the alternative splicing variant of cyclin D1. PACSIN 2 knockdown induced cellular migration and reduced cell spreading in LNCaP cells expressing cyclin D1a. In cyclin D1-/- mouse embryonic fibroblasts (MeFs), cyclin D1a, but not cyclin D1b, reduced the cell spreading to a polarized morphology. sipACSIN 2 had no effect on cellular migration of cyclin D1 -/- MeFs. Cyclin D1a restored the migratory ability of cyclin D1 -/- MeFs, which was further enhanced by knocking down PACSIN 2 with siRNA. the cyclin D1-associated protein, PACSIN 2, regulates cell spreading and migration, which are dependent on cyclin D1 expression.
UR - http://www.scopus.com/inward/record.url?scp=78650833036&partnerID=8YFLogxK
U2 - 10.4161/cc.10.1.14243
DO - 10.4161/cc.10.1.14243
M3 - Article
C2 - 21200149
AN - SCOPUS:78650833036
SN - 1538-4101
VL - 10
SP - 73
EP - 81
JO - Cell Cycle
JF - Cell Cycle
IS - 1
ER -