Concerted effects of heterogeneous nuclear ribonucleoprotein C1/C2 to control vitamin D-directed gene transcription and RNA splicing in human bone cells.
Traditionally recognized as an RNA splicing regulator, heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNPC1/C2) can also bind to double-stranded DNA and function in trans as a vitamin D response element (VDRE)-binding protein. As such, hnRNPC1/C2 may couple transcription induced by the active form of vitamin D, 1,25-dihydroxyvitamin D (1,25(OH)2D) with subsequent RNA splicing. In MG63 osteoblastic cells, increased expression of the 1,25(OH)2D target gene CYP24A1 involved immunoprecipitation of hnRNPC1/C2 with CYP24A1 chromatin and RNA. Knockdown of hnRNPC1/C2 suppressed expression of CYP24A1, but also increased expression of an exon 10-skipped CYP24A1 splice variant; in a minigene model the latter was attenuated by a functional VDRE in the CYP24A1 promoter. In genome-wide analyses, knockdown of hnRNPC1/C2 resulted in 3500 differentially expressed genes and 2232 differentially spliced genes, with significant commonality between groups. 1,25(OH)2D induced 324 differentially expressed genes, with 187 also observed following hnRNPC1/C2 knockdown, and a further 168 unique to hnRNPC1/C2 knockdown. However, 1,25(OH)2D induced only 10 differentially spliced genes, with no overlap with differentially expressed genes. These data indicate that hnRNPC1/C2 binds to both DNA and RNA and influences both gene expression and RNA splicing, but these actions do not appear to be linked through 1,25(OH)2D-mediated induction of transcription.
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.|Department of Orthopaedics, the Orthopedic Surgery Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China.
Rui
R
Zhou
Microbiology, Immunology and Molecular Genetics, University of California at Los Angeles, Los Angeles, CA 90095, USA.|Computer Engineering and Computer Science, Kentucky Biomedical Research Infrastructure Network, Louisville, KY 40292, USA.
Juw Won
JW
Park
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Rene F
RF
Chun
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
Thomas S
TS
Lisse
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Alejandro J
AJ
Garcia
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Kathryn
K
Zavala
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Jessica L
JL
Sea
Microbiology, Immunology and Molecular Genetics, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Zhi-Xiang
ZX
Lu
Department of Orthopaedics, the Orthopedic Surgery Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China.
Jianzhong
J
Xu
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA.
John S
JS
Adams
Microbiology, Immunology and Molecular Genetics, University of California at Los Angeles, Los Angeles, CA 90095, USA yxing@ucla.edu.
Yi
Y
Xing
UCLA Orthopaedic Hospital, Department of Orthopaedic Surgery, Orthopedic Hospital, University of California at Los Angeles, Los Angeles, CA 90095, USA m.hewison@bham.ac.uk.|Institute of Metabolism and Systems Research, the University of Birmingham, Birmingham, B15 2TT, UK.
Martin
0000-0001-5806-9690
M
Hewison
D017398:Alternative Splicing; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C / Q000235:genetics / Q000378:metabolism*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D010011:Osteocytes / Q000187:drug effects / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034622:RNA Interference; D012322:RNA Precursors; D012326:RNA Splicing; D014158:Transcription, Genetic; D014807:Vitamin D / Q000378:metabolism*; D065668:Vitamin D3 24-Hydroxylase / Q000235:genetics / Q000378:metabolism
D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D012322:RNA Precursors; D014807:Vitamin D; D065668:Vitamin D3 24-Hydroxylase
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