Analysis of PTEN complex assembly and identification of heterogeneous nuclear ribonucleoprotein C as a component of the PTEN-associated complex.
PTEN (phosphatase and tensin homolog deleted on chromosome 10) is well characterized for its role in antagonizing the phosphoinositide 3-kinase pathway. Previous studies using size-exclusion chromatography demonstrated PTEN recruitment into high molecular mass complexes and hypothesized that PTEN phosphorylation status and PDZ binding domain may be required for such complex formation. In this study, we set out to test the structural requirements for PTEN complex assembly and identify the component(s) of the PTEN complex(es). Our results demonstrated that the PTEN catalytic function and PDZ binding domain are not absolutely required for its complex formation. On the other hand, PTEN phosphorylation status has a significant impact on its complex assembly. Our results further demonstrate enrichment of the PTEN complex in nuclear lysates, suggesting a mechanism through which PTEN phosphorylation may regulate its complex assembly. These results prompted further characterization of other protein components within the PTEN complex(es). Using size-exclusion chromatography and two-dimensional difference gel electrophoresis followed by mass spectrometry analysis, we identified heterogeneous nuclear ribonucleoprotein C (hnRNP C) as a novel protein recruited to higher molecular mass fractions in the presence of PTEN. Further analysis indicates that endogenous hnRNP C and PTEN interact and co-localize within the nucleus, suggesting a potential role for PTEN, alongside hnRNP C, in RNA regulation.
Mosessian
Sherly
S
Department of Molecular and Medical Pharmacology, Institute for Molecular Medicine, University of California, Los Angeles, California 90095, USA.
Avliyakulov
Nuraly K
NK
Mulholland
David J
DJ
Boontheung
Pinmanee
P
Loo
Joseph A
JA
Wu
Hong
H
D021581:Active Transport, Cell Nucleus; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D046912:Multiprotein Complexes / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism; D051059:PTEN Phosphohydrolase / Q000378:metabolism*; D010766:Phosphorylation
D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human