PMID- 26845344 OWN - NLM STAT- MEDLINE DCOM- 20170925 LR - 20220321 IS - 1872-8057 (Electronic) IS - 0303-7207 (Linking) VI - 430 DP - 2016 Jul 15 TI - HMGB1 binds to the rs7903146 locus in TCF7L2 in human pancreatic islets. PG - 138-45 LID - S0303-7207(16)30027-2 [pii] LID - 10.1016/j.mce.2016.01.027 [doi] AB - The intronic SNP rs7903146 in the T-cell factor 7-like 2 gene (TCF7L2) is the common genetic variant most highly associated with Type 2 diabetes known to date. The risk T-allele is located in an open chromatin region specific to human pancreatic islets of Langerhans, thereby accessible for binding of regulatory proteins. The risk T-allele locus exhibits stronger enhancer activity compared to the non-risk C-allele. The aim of this study was to identify transcriptional regulators that bind the open chromatin region in the rs7903146 locus and thereby potentially regulate TCF7L2 expression and activity. Using affinity chromatography followed by Edman sequencing, we identified one candidate regulatory protein, i.e. high-mobility group protein B1 (HMGB1). The binding of HMGB1 to the rs7903146 locus was confirmed in pancreatic islets from human deceased donors, in HCT116 and in HEK293 cell lines using: (i) protein purification on affinity columns followed by Western blot, (ii) chromatin immunoprecipitation followed by qPCR and (iii) electrophoretic mobility shift assay. The results also suggested that HMGB1 might have higher binding affinity to the C-allele of rs7903146 compared to the T-allele, which was supported in vitro using Dynamic Light Scattering, possibly in a tissue-specific manner. The functional consequence of HMGB1 depletion in HCT116 and INS1 cells was reduced insulin and TCF7L2 mRNA expression, TCF7L2 transcriptional activity and glucose stimulated insulin secretion. These findings suggest that the rs7903146 locus might exert its enhancer function by interacting with HMGB1 in an allele dependent manner. CI - Copyright (c) 2016 Elsevier Ireland Ltd. All rights reserved. FAU - Zhou, Yuedan AU - Zhou Y AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Oskolkov, Nikolay AU - Oskolkov N AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Shcherbina, Liliya AU - Shcherbina L AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Ratti, Joyce AU - Ratti J AD - Department of Biochemistry, University of Cambridge, CB2 1GA, Cambridge, UK. FAU - Kock, Kian-Hong AU - Kock KH AD - Department of Biochemistry, University of Cambridge, CB2 1GA, Cambridge, UK. FAU - Su, Jing AU - Su J AD - European Bioinformatics Institute, Functional Genomics, Hinxton, Cambridge CB10 1SD, UK. FAU - Martin, Brian AU - Martin B AD - National Institute of Mental Health NIMH, National Institutes of Health, Bethesda, MD 20892, USA. FAU - Oskolkova, Malin Zackrisson AU - Oskolkova MZ AD - Department of Physical Chemistry, Lund University, Lund, 22100, Sweden. FAU - Goransson, Olga AU - Goransson O AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Bacon, Julie AU - Bacon J AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Li, Weimin AU - Li W AD - Department of Physical Chemistry, Lund University, Lund, 22100, Sweden. FAU - Bucciarelli, Saskia AU - Bucciarelli S AD - Department of Physical Chemistry, Lund University, Lund, 22100, Sweden. FAU - Cilio, Corrado AU - Cilio C AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Brazma, Alvis AU - Brazma A AD - European Bioinformatics Institute, Functional Genomics, Hinxton, Cambridge CB10 1SD, UK. FAU - Thatcher, Bradley AU - Thatcher B AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Rung, Johan AU - Rung J AD - European Bioinformatics Institute, Functional Genomics, Hinxton, Cambridge CB10 1SD, UK. FAU - Wierup, Nils AU - Wierup N AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Renstrom, Erik AU - Renstrom E AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Groop, Leif AU - Groop L AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. FAU - Hansson, Ola AU - Hansson O AD - Department of Clinical Sciences, CRC, Lund University, Malmo, 20502, Sweden. Electronic address: Ola.Hansson@med.lu.se. LA - eng GR - Z01 MH002344/MH/NIMH NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Intramural PT - Research Support, Non-U.S. Gov't DEP - 20160201 PL - Ireland TA - Mol Cell Endocrinol JT - Molecular and cellular endocrinology JID - 7500844 RN - 0 (HMGB1 Protein) RN - 0 (RNA, Messenger) RN - 0 (TCF7L2 protein, human) RN - 0 (Transcription Factor 7-Like 2 Protein) RN - 9007-49-2 (DNA) SB - IM MH - Animals MH - Computer Simulation MH - DNA/metabolism MH - Dynamic Light Scattering MH - *Genetic Loci MH - HCT116 Cells MH - HEK293 Cells MH - HMGB1 Protein/*metabolism MH - Humans MH - Hydrodynamics MH - Islets of Langerhans/*metabolism MH - Polymorphism, Single Nucleotide/*genetics MH - Protein Binding MH - RNA, Messenger/genetics/metabolism MH - Rats MH - Reproducibility of Results MH - Transcription Factor 7-Like 2 Protein/*genetics OTO - NOTNLM OT - Chromatin binding OT - HMGB1 OT - TCF7L2 OT - Type 2 diabetes EDAT- 2016/02/05 06:00 MHDA- 2017/09/26 06:00 CRDT- 2016/02/05 06:00 PHST- 2015/10/02 00:00 [received] PHST- 2016/01/19 00:00 [revised] PHST- 2016/01/28 00:00 [accepted] PHST- 2016/02/05 06:00 [entrez] PHST- 2016/02/05 06:00 [pubmed] PHST- 2017/09/26 06:00 [medline] AID - S0303-7207(16)30027-2 [pii] AID - 10.1016/j.mce.2016.01.027 [doi] PST - ppublish SO - Mol Cell Endocrinol. 2016 Jul 15;430:138-45. doi: 10.1016/j.mce.2016.01.027. Epub 2016 Feb 1.