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Effect of APOL1 disease risk variants on APOL1 gene product
Gene sequence mutations may alter mRNA transcription, transcript stability, protein translation, protein stability and protein folding. Apolipoprotein L1 (APOL1) has two sets of sequence variants that are risk factors for kidney disease development, APOL1G1 (substitution mutation) and APOL1G2 (delet...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408699/ https://www.ncbi.nlm.nih.gov/pubmed/28385815 http://dx.doi.org/10.1042/BSR20160531 |
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author | Haque, Shabirul Patil, Gauri Mishra, Abheepsa Lan, Xiqian Popik, Waldemar Malhotra, Ashwani Skorecki, Karl Singhal, Pravin C. |
author_facet | Haque, Shabirul Patil, Gauri Mishra, Abheepsa Lan, Xiqian Popik, Waldemar Malhotra, Ashwani Skorecki, Karl Singhal, Pravin C. |
author_sort | Haque, Shabirul |
collection | PubMed |
description | Gene sequence mutations may alter mRNA transcription, transcript stability, protein translation, protein stability and protein folding. Apolipoprotein L1 (APOL1) has two sets of sequence variants that are risk factors for kidney disease development, APOL1G1 (substitution mutation) and APOL1G2 (deletion mutation). Our present study focuses on the impact of these variants on APOL1 mRNA transcription and translation. APOL1 plasmids (EV, G0, G1 and G2) were transfected into human embryonic kidney (HEK) 293T cells. APOL1 variant expression was observed to be significantly lower than that of APOL1G0. Podocyte cell lines stably expressing APOL1 transgenes also showed lower levels of APOL1 expression of APOL1 variants (G1 and G2) compared with APOL1G0 by Western blotting and FACS analysis. The enhanced expression of GRP78 by podocytes expressing APOL1 variants would indicate endoplasmic reticulum (ER) stress. Bioinformatics evaluation using two different programs (MUPro and I-Mutant 2.0) predicted that APOL1 variants were less stable than APOL1G0. Concomitant with protein levels, APOL1 mRNA levels were also depressed following induction of APOL1 variant compared with APOL1G0 in both proliferating and differentiated podocytes. APOL1 mRNA transcript stability was tested after actinomycin D pulsing; APOL1G1 and APOL1G2 mRNAs transcript decayed 10–15% and 15–20% (within a period of 0.5–3 h) respectively. Our data suggest that down-regulated APOL1 protein expression in APOL1 variants is due to compromised transcription and decay of the APOL1 variant transcripts. |
format | Online Article Text |
id | pubmed-5408699 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-54086992017-05-04 Effect of APOL1 disease risk variants on APOL1 gene product Haque, Shabirul Patil, Gauri Mishra, Abheepsa Lan, Xiqian Popik, Waldemar Malhotra, Ashwani Skorecki, Karl Singhal, Pravin C. Biosci Rep Research Articles Gene sequence mutations may alter mRNA transcription, transcript stability, protein translation, protein stability and protein folding. Apolipoprotein L1 (APOL1) has two sets of sequence variants that are risk factors for kidney disease development, APOL1G1 (substitution mutation) and APOL1G2 (deletion mutation). Our present study focuses on the impact of these variants on APOL1 mRNA transcription and translation. APOL1 plasmids (EV, G0, G1 and G2) were transfected into human embryonic kidney (HEK) 293T cells. APOL1 variant expression was observed to be significantly lower than that of APOL1G0. Podocyte cell lines stably expressing APOL1 transgenes also showed lower levels of APOL1 expression of APOL1 variants (G1 and G2) compared with APOL1G0 by Western blotting and FACS analysis. The enhanced expression of GRP78 by podocytes expressing APOL1 variants would indicate endoplasmic reticulum (ER) stress. Bioinformatics evaluation using two different programs (MUPro and I-Mutant 2.0) predicted that APOL1 variants were less stable than APOL1G0. Concomitant with protein levels, APOL1 mRNA levels were also depressed following induction of APOL1 variant compared with APOL1G0 in both proliferating and differentiated podocytes. APOL1 mRNA transcript stability was tested after actinomycin D pulsing; APOL1G1 and APOL1G2 mRNAs transcript decayed 10–15% and 15–20% (within a period of 0.5–3 h) respectively. Our data suggest that down-regulated APOL1 protein expression in APOL1 variants is due to compromised transcription and decay of the APOL1 variant transcripts. Portland Press Ltd. 2017-04-28 /pmc/articles/PMC5408699/ /pubmed/28385815 http://dx.doi.org/10.1042/BSR20160531 Text en © 2017 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Articles Haque, Shabirul Patil, Gauri Mishra, Abheepsa Lan, Xiqian Popik, Waldemar Malhotra, Ashwani Skorecki, Karl Singhal, Pravin C. Effect of APOL1 disease risk variants on APOL1 gene product |
title | Effect of APOL1 disease risk variants on APOL1 gene product |
title_full | Effect of APOL1 disease risk variants on APOL1 gene product |
title_fullStr | Effect of APOL1 disease risk variants on APOL1 gene product |
title_full_unstemmed | Effect of APOL1 disease risk variants on APOL1 gene product |
title_short | Effect of APOL1 disease risk variants on APOL1 gene product |
title_sort | effect of apol1 disease risk variants on apol1 gene product |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408699/ https://www.ncbi.nlm.nih.gov/pubmed/28385815 http://dx.doi.org/10.1042/BSR20160531 |
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