Alternative Splicing Regulation of Caspase-2 Gene

Project: Research project

Project Details

Description

DESCRIPTION (provided by applicant): Pre-mRNA splicing is a critical step in the expression of most mammalian genes. Alternative splicing is a fundamental mechanism that contributes to genetic diversity. Our long-term goal is to understand the molecular basis underlying the regulation of mammalian pre-mRNA splicing and its role in the pathogenesis and treatment of cancer. This application focuses on investigating mechanisms underlying splice site recognition/selection and the role of alternative splicing regulation of caspase-2 (casp-2), an important player in programmed cell death (PCD). A number of important PCD genes, including casp-2, utilize alternative splicing to generate functionally antagonistic products. Recent studies suggest that casp-2 is critical for the initiation of cell death induced by anti-cancer drugs. Our preliminary results show that alternative splicing pattern of casp-2 changes in response to chemotherapeutic drugs. To investigate mechanisms underlying the alternative splicing regulation of caspase genes, we have established a model system using casp-2 gene. A casp-2 minigene has been constructed containing cis-elements essential and sufficient for reproducing casp-2 alternative splicing. Casp-2 alternative splicing has been reconstituted in vitro. Using both in vitro biochemical assays and transfection in the cultured cells, we have begun to dissect cis-elements and transacting factors critical for casp-2 alternative splicing. We propose to study molecular mechanisms regulating casp-2 alternative splicing by dissecting cis- elements and trans-acting factors involved. We plan to use combined molecular and biochemical approaches to investigate how different cis- and trans-factors function in regulating casp-2 alternative splicing that controls the delicate balance of anti- and pro- apoptotic products. We will use targeted-knock-out approach to study the role of In100 in regulating casp-2 alternative splicing in mice. We will examine changes in PCD gene alternative splicing and in the expression/activities of splicing regulators during cell death. This study is likely to provide new insights into mechanisms of mammalian splicing regulation and to advance our understanding of regulation of PCD.
StatusFinished
Effective start/end date3/1/062/28/09

Funding

  • National Institute of General Medical Sciences (5 R01 GM070967-03)

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