Requirement for High Mobility Group Protein HMGI-C Interaction with STAT3 Inhibitor PIAS3 in Repression of α-Subunit of Epithelial Na+Channel (α-ENaC) Transcription by Ras Activation in Salivary Epithelial Cells

Mark D. Zentner, H. Helen Lin, Hong-Tao Deng, Kwang-Jin Kim, Hsiu-Ming Shih, David K. Ann
2001 Journal of Biological Chemistry  
Previously, we have demonstrated that oxidative stress or Ras/ERK activation leads to the transcriptional repression of ␣-subunit of epithelial Na ؉ channel (ENaC) in lung and salivary epithelial cells. Here, we further investigated the coordinated molecular mechanisms by which ␣-ENaC expression is regulated. Using both stable and transient transfection assays, we demonstrate that the overexpression of high mobility group protein I-C (HMGI-C), a Ras/ERK-inducible HMG-I family member, represses
more » ... member, represses glucocorticoid receptor (GR)/dexamethasone (Dex)-stimulated ␣-ENaC/reporter activity in salivary epithelial cells. Northern analyses further confirm that the expression of endogenous ␣-ENaC gene in salivary Pa-4 cells is suppressed by an ectopic HMGI-C overexpression. Through yeast two-hybrid screening and co-immunoprecipitation assays from eukaryotic cells, we also discovered the interaction between HMGI-C and PIAS3 (protein inhibitor of activated STAT3 (signal transducer and activator of transcription 3)). A low level of ectopically expressed PIAS3 cooperatively inhibits GR/Dex-dependent ␣-ENaC transcription in the presence of HMGI-C. Reciprocally, HMGI-C expression also coordinately enhances PIAS3-mediated repression of STAT3-dependent transactivation. Moreover, overexpression of antisense HMGI-C construct is capable of reversing the repression mediated by Ras V12 on GR-and STAT3-dependent transcriptional activation. Together, our results demonstrate that Ras/ERKmediated induction of HMGI-C is required to effectively repress GR/Dex-stimulated transcription of ␣-ENaC gene and STAT3-mediated transactivation. These findings delineate a network of inhibitory signaling pathways that converge on HMGI-C⅐PIAS3 complex, causally associating Ras/ERK activation with the repression of both GR and STAT3 signaling pathways in salivary epithelial cells.
doi:10.1074/jbc.m103153200 pmid:11390395 fatcat:wafwfvw2izgdzmcqz5l2jn5cni