Cargando…

Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection

Promyelocytic leukemia (PML) protein is a crucial component of PML-nuclear bodies (PML-NBs). PML and PML-NBs are involved in the regulation of various cellular functions, including the antiviral immune response. The human PML gene can generate several different isoforms through alternative splicing....

Descripción completa

Detalles Bibliográficos
Autores principales: Zhu, Jingjing, Chen, Zhenyu, Dai, Zhenglie, Zhou, Xiaolong, Wang, Han, Li, Xiangchen, Zhao, Ayong, Yang, Songbai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8649775/
https://www.ncbi.nlm.nih.gov/pubmed/34888375
http://dx.doi.org/10.3389/fvets.2021.757978
_version_ 1784611070721130496
author Zhu, Jingjing
Chen, Zhenyu
Dai, Zhenglie
Zhou, Xiaolong
Wang, Han
Li, Xiangchen
Zhao, Ayong
Yang, Songbai
author_facet Zhu, Jingjing
Chen, Zhenyu
Dai, Zhenglie
Zhou, Xiaolong
Wang, Han
Li, Xiangchen
Zhao, Ayong
Yang, Songbai
author_sort Zhu, Jingjing
collection PubMed
description Promyelocytic leukemia (PML) protein is a crucial component of PML-nuclear bodies (PML-NBs). PML and PML-NBs are involved in the regulation of various cellular functions, including the antiviral immune response. The human PML gene can generate several different isoforms through alternative splicing. However, little is known about the porcine PML alternative splicing isoforms and their expression profiles during Japanese encephalitis virus (JEV) infection. In the present study, we cloned seven mature transcripts of porcine PML, all of which contained the same N-terminal sequence but differed in the C-terminal sequences due to alternative splicing. These seven transcripts encoded five proteins all of which had the RBCC motif and sumoylation sites. Amino acid sequence homology analysis showed that porcine PML-1 had relatively high levels of identity with human, cattle, and goat homologs (76.21, 77.17, and 77.05%, respectively), and low identity with the mouse homolog (61.78%). Immunofluorescence analysis showed that the typical PML-NBs could be observed after overexpression of the five PML isoforms in PK15 cells. Quantitative reverse transcription PCR (RT-qPCR) analysis showed significant upregulation of PML isoforms and PML-NB-associated genes (Daxx and SP100) at 36 and 48 h post-infection (hpi). Western blotting analysis indicated that the PML isoforms were upregulated during the late stage of infection. Moreover, the number of PML-NBs was increased after JEV infection. These results suggest that porcine PML isoforms may play essential roles in JEV infection.
format Online
Article
Text
id pubmed-8649775
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-86497752021-12-08 Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection Zhu, Jingjing Chen, Zhenyu Dai, Zhenglie Zhou, Xiaolong Wang, Han Li, Xiangchen Zhao, Ayong Yang, Songbai Front Vet Sci Veterinary Science Promyelocytic leukemia (PML) protein is a crucial component of PML-nuclear bodies (PML-NBs). PML and PML-NBs are involved in the regulation of various cellular functions, including the antiviral immune response. The human PML gene can generate several different isoforms through alternative splicing. However, little is known about the porcine PML alternative splicing isoforms and their expression profiles during Japanese encephalitis virus (JEV) infection. In the present study, we cloned seven mature transcripts of porcine PML, all of which contained the same N-terminal sequence but differed in the C-terminal sequences due to alternative splicing. These seven transcripts encoded five proteins all of which had the RBCC motif and sumoylation sites. Amino acid sequence homology analysis showed that porcine PML-1 had relatively high levels of identity with human, cattle, and goat homologs (76.21, 77.17, and 77.05%, respectively), and low identity with the mouse homolog (61.78%). Immunofluorescence analysis showed that the typical PML-NBs could be observed after overexpression of the five PML isoforms in PK15 cells. Quantitative reverse transcription PCR (RT-qPCR) analysis showed significant upregulation of PML isoforms and PML-NB-associated genes (Daxx and SP100) at 36 and 48 h post-infection (hpi). Western blotting analysis indicated that the PML isoforms were upregulated during the late stage of infection. Moreover, the number of PML-NBs was increased after JEV infection. These results suggest that porcine PML isoforms may play essential roles in JEV infection. Frontiers Media S.A. 2021-11-23 /pmc/articles/PMC8649775/ /pubmed/34888375 http://dx.doi.org/10.3389/fvets.2021.757978 Text en Copyright © 2021 Zhu, Chen, Dai, Zhou, Wang, Li, Zhao and Yang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Zhu, Jingjing
Chen, Zhenyu
Dai, Zhenglie
Zhou, Xiaolong
Wang, Han
Li, Xiangchen
Zhao, Ayong
Yang, Songbai
Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title_full Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title_fullStr Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title_full_unstemmed Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title_short Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
title_sort molecular cloning of alternative splicing variants of the porcine pml gene and its expression patterns during japanese encephalitis virus infection
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8649775/
https://www.ncbi.nlm.nih.gov/pubmed/34888375
http://dx.doi.org/10.3389/fvets.2021.757978
work_keys_str_mv AT zhujingjing molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT chenzhenyu molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT daizhenglie molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT zhouxiaolong molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT wanghan molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT lixiangchen molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT zhaoayong molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection
AT yangsongbai molecularcloningofalternativesplicingvariantsoftheporcinepmlgeneanditsexpressionpatternsduringjapaneseencephalitisvirusinfection