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Eur J Biochem ; 271(19): 3812-20, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15373827

RESUMO

HIP/PAP is a C-type lectin overexpressed in hepatocellular carcinoma (HCC). Pleiotropic biological activities have been ascribed to this protein, but little is known about the function of HIP/PAP in the liver. In this study, therefore, we searched for proteins interacting with HIP/PAP by screening a HCC cDNA expression library. We have identified the RII alpha regulatory subunit of cAMP-dependent protein kinase (PKA) as a partner of HIP/PAP. HIP/PAP and RII alpha were coimmunoprecipitated in HIP/PAP expressing cells. The biological relevance of the interaction between these proteins was established by demonstrating, using fractionation methods, that they are located in a same subcellular compartment. Indeed, though HIP/PAP is a protein secreted via the Golgi apparatus we showed that a fraction of HIP/PAP escaped the secretory apparatus and was recovered in the cytosol. Basal PKA activity was increased in HIP/PAP expressing cells, suggesting that HIP/PAP may alter PKA signalling. Indeed, we showed, using a thymidine kinase-luciferase reporter plasmid in which a cAMP responsive element was inserted upstream of the thymidine kinase promoter, that luciferase activity was enhanced in HIP/PAP expressing cells. Thus our findings suggest a novel mechanism for the biological activity of the HIP/PAP lectin.


Assuntos
Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Lectinas Tipo C/metabolismo , Neoplasias Hepáticas/metabolismo , Transdução de Sinais , Antígenos de Neoplasias/genética , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Subunidade RIIbeta da Proteína Quinase Dependente de AMP Cíclico , Biblioteca Gênica , Complexo de Golgi , Humanos , Imunoprecipitação , Lectinas Tipo C/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Luciferases/metabolismo , Proteínas Associadas a Pancreatite , Fosforilação , Frações Subcelulares , Timidina Quinase/metabolismo , Células Tumorais Cultivadas
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