The research plan is divided into two parts, regulation at the transcriptional level and regulation at the translational level. The first part of the project is the continuation of studies to characterize the proteins that are responsible for the induction of connexin43 in the myometrium. The functional characterization of one of these proteins, a novel transcription factor, Ini, that was discovered during the previous grant period, will be completed. Three additional transcription factors that were identified by Southwestern blot and DNA mobility shift analysis will be cloned and the function of these proteins in connexin43 gene regulation studied. The likely presence of an additional estrogen-responsive transcriptional activation site in the 5'-UTR of the connexin43 gene will also be investigated. The second part of this research is based on the recent discovery in this laboratory of an internal ribosome entry site (IRES) in the 5'-UTR of the connexin43 mRNA. The function of the IRES element will be studied in bicistroni vector constructs both in vivo and in vitro. In addition, the proteins that function in the activation of the connexin43 IRES will be identified by RNA mobility shift analysis and ribosome binding assays.
|Anderson, Curtis; Catoe, Heath; Werner, Rudolf (2006) MIR-206 regulates connexin43 expression during skeletal muscle development. Nucleic Acids Res 34:5863-71|
|Anderson, Curtis L; Zundel, Michael A; Werner, Rudolf (2005) Variable promoter usage and alternative splicing in five mouse connexin genes. Genomics 85:238-44|
|Pfeifer, Ingrid; Anderson, Curtis; Werner, Rudolf et al. (2004) Redefining the structure of the mouse connexin43 gene: selective promoter usage and alternative splicing mechanisms yield transcripts with different translational efficiencies. Nucleic Acids Res 32:4550-62|
|Oltra, Elisa; Verde, Fulvia; Werner, Rudolf et al. (2004) A novel RING-finger-like protein Ini1 is essential for cell cycle progression in fission yeast. J Cell Sci 117:967-74|
|Oltra, Elisa; Pfeifer, Ingrid; Werner, Rudolf (2003) Ini, a small nuclear protein that enhances the response of the connexin43 gene to estrogen. Endocrinology 144:3148-58|
|Hudder, A; Werner, R (2000) Analysis of a Charcot-Marie-Tooth disease mutation reveals an essential internal ribosome entry site element in the connexin-32 gene. J Biol Chem 275:34586-91|
|Schiavi, A; Hudder, A; Werner, R (1999) Connexin43 mRNA contains a functional internal ribosome entry site. FEBS Lett 464:118-22|