title: Stimulation of StAR expression by cAMP is controlled by inhibition of highly inducible SIK1 via CRTC2, a co-activator of CREB.
abstract: In mouse steroidogenic cells the activation of cholesterol metabolism is mediated by steroidogenic acute regulatory protein (StAR). Here, we visualized a coordinated regulation of StAR transcription, splicing and post-transcriptional processing, which are synchronized by salt inducible kinase (SIK1) and CREB-regulated transcription coactivator (CRTC2). To detect primary RNA (pRNA), spliced primary RNA (Sp-RNA) and mRNA in single cells, we generated probe sets by using fluorescence in situ hybridization (FISH). These methods allowed us to address the nature of StAR gene expression and to visualize protein-nucleic acid interactions through direct detection. We show that SIK1 represses StAR expression in Y1 adrenal and MA10 testis cells through inhibition of processing mediated by CRTC2. Digital image analysis matches qPCR analyses of the total cell culture. Evidence is presented for spatially separate accumulation of StAR pRNA and Sp-RNA at the gene loci in the nucleus. These findings establish that cAMP, SIK and CRTC mediate StAR expression through activation of individual StAR gene loci.
authors: Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI, USA; Endocrinology and Reproductive Physiology Program, University of Wisconsin, Madison, WI, USA.
authors: Jinwoo
authors: J
authors: Lee
authors: Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI, USA.
authors: Tiegang
authors: T
authors: Tong
authors: National Institute for Biomedical Innovation, Osaka, Japan.
authors: Hiroshi
authors: H
authors: Takemori
authors: Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI, USA; Endocrinology and Reproductive Physiology Program, University of Wisconsin, Madison, WI, USA; University of Wisconsin School of Medicine and Public Health, Madison, WI, USA. Electronic address: jefcoate@wisc.edu.
authors: Colin
authors: C
authors: Jefcoate
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D017398:Alternative Splicing / Q000187:drug effects / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004790:Enzyme Induction / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008954:Models, Biological; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D012322:RNA Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: C504923:Crtc2 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; D010750:Phosphoproteins; D012322:RNA Precursors; D012333:RNA, Messenger; D014157:Transcription Factors; D000098682:Steroidogenic Acute Regulatory Protein; D049452:Green Fluorescent Proteins; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP; D017346:Protein Serine-Threonine Kinases; C495376:Sik1 protein, mouse
keywords: CRTC; Fluorescence in situ hybridization; SIK; Splicing; StAR