ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition

Human ZAP inhibits many viruses, including HIV and coronaviruses, by binding to viral RNAs to promote their degradation and/or translation suppression. However, the regulatory role of ZAP in host mRNAs is largely unknown. Two major alternatively spliced ZAP isoforms, the constitutively expressed ZAP...

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Main Authors: Ly, Phuong Thao, Xu, Shaohai, Wirawan, Melissa, Luo, Dahai, Roca, Xavier
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2023
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Online Access:https://hdl.handle.net/10356/164082
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spelling sg-ntu-dr.10356-1640822023-02-28T16:57:08Z ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition Ly, Phuong Thao Xu, Shaohai Wirawan, Melissa Luo, Dahai Roca, Xavier School of Biological Sciences Lee Kong Chian School of Medicine (LKCMedicine) Science::Biological sciences Alternative Splicing Unfolded Protein Response Human ZAP inhibits many viruses, including HIV and coronaviruses, by binding to viral RNAs to promote their degradation and/or translation suppression. However, the regulatory role of ZAP in host mRNAs is largely unknown. Two major alternatively spliced ZAP isoforms, the constitutively expressed ZAPL and the infection-inducible ZAPS, play overlapping yet different antiviral and other roles that need further characterization. We found that the splicing factors hnRNPA1/A2, PTBP1/2, and U1-snRNP inhibit ZAPS production and demonstrated the feasibility to modulate the ZAPL/S balance by splice-switching antisense oligonucleotides in human cells. Transcriptomic analysis of ZAP-isoform-specific knockout cells revealed uncharacterized host mRNAs targeted by ZAPL/S with broad cellular functions such as unfolded protein response (UPR), epithelial-mesenchymal transition (EMT), and innate immunity. We established that endogenous ZAPL and ZAPS localize to membrane compartments and cytosol, respectively, and that the differential localization correlates with their target-RNA specificity. We showed that the ZAP isoforms regulated different UPR branches under resting and stress conditions and affected cell viability during ER stress. We also provided evidence for a different function of the ZAP isoforms in EMT-related cell migration, with effects that are cell-type dependent. Overall, this study demonstrates that the competition between splicing and IPA is a potential target for the modulation of the ZAPL/S balance, and reports new cellular transcripts and processes regulated by the ZAP isoforms. Ministry of Education (MOE) Nanyang Technological University National Medical Research Council (NMRC) National Research Foundation (NRF) Published version This work was funded by the NTU Integrated Medical Biological and Environmental Life Sciences NIM/02/2017 (to X.R. and D.L.), Academic Research Fund Tier 1 (RG22/21) from Singapore’s Ministry of Education (to X.R.), NRF2019-NRF-ISF003-3104 from the National Research Foundation of Singapore (to X.R.), and the OFIRG17nov084 grant from Singapore’s National Medical Research Council (to D.L.). 2023-01-04T05:13:35Z 2023-01-04T05:13:35Z 2022 Journal Article Ly, P. T., Xu, S., Wirawan, M., Luo, D. & Roca, X. (2022). ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition. Proceedings of the National Academy of Sciences of the United States of America, 119(31), e2121453119-. https://dx.doi.org/10.1073/pnas.2121453119 0027-8424 https://hdl.handle.net/10356/164082 10.1073/pnas.2121453119 35881805 2-s2.0-85135102366 31 119 e2121453119 en NIM/02/2017 RG22/21 NRF2019-NRF-ISF003-3104 OFIRG17nov084 Proceedings of the National Academy of Sciences of the United States of America © 2022 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND). application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
Alternative Splicing
Unfolded Protein Response
spellingShingle Science::Biological sciences
Alternative Splicing
Unfolded Protein Response
Ly, Phuong Thao
Xu, Shaohai
Wirawan, Melissa
Luo, Dahai
Roca, Xavier
ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
description Human ZAP inhibits many viruses, including HIV and coronaviruses, by binding to viral RNAs to promote their degradation and/or translation suppression. However, the regulatory role of ZAP in host mRNAs is largely unknown. Two major alternatively spliced ZAP isoforms, the constitutively expressed ZAPL and the infection-inducible ZAPS, play overlapping yet different antiviral and other roles that need further characterization. We found that the splicing factors hnRNPA1/A2, PTBP1/2, and U1-snRNP inhibit ZAPS production and demonstrated the feasibility to modulate the ZAPL/S balance by splice-switching antisense oligonucleotides in human cells. Transcriptomic analysis of ZAP-isoform-specific knockout cells revealed uncharacterized host mRNAs targeted by ZAPL/S with broad cellular functions such as unfolded protein response (UPR), epithelial-mesenchymal transition (EMT), and innate immunity. We established that endogenous ZAPL and ZAPS localize to membrane compartments and cytosol, respectively, and that the differential localization correlates with their target-RNA specificity. We showed that the ZAP isoforms regulated different UPR branches under resting and stress conditions and affected cell viability during ER stress. We also provided evidence for a different function of the ZAP isoforms in EMT-related cell migration, with effects that are cell-type dependent. Overall, this study demonstrates that the competition between splicing and IPA is a potential target for the modulation of the ZAPL/S balance, and reports new cellular transcripts and processes regulated by the ZAP isoforms.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Ly, Phuong Thao
Xu, Shaohai
Wirawan, Melissa
Luo, Dahai
Roca, Xavier
format Article
author Ly, Phuong Thao
Xu, Shaohai
Wirawan, Melissa
Luo, Dahai
Roca, Xavier
author_sort Ly, Phuong Thao
title ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
title_short ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
title_full ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
title_fullStr ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
title_full_unstemmed ZAP isoforms regulate unfolded protein response and epithelial- mesenchymal transition
title_sort zap isoforms regulate unfolded protein response and epithelial- mesenchymal transition
publishDate 2023
url https://hdl.handle.net/10356/164082
_version_ 1759858298865057792