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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...

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Autores principales: Haque, Shabirul, Patil, Gauri, Mishra, Abheepsa, Lan, Xiqian, Popik, Waldemar, Malhotra, Ashwani, Skorecki, Karl, Singhal, Pravin C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2017
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.
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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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