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....

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Autores principales:	Jingjing Zhu, Zhenyu Chen, Zhenglie Dai, Xiaolong Zhou, Han Wang, Xiangchen Li, Ayong Zhao, Songbai Yang
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	porcine PML gene alternative splicing clone JEV Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/3974c584ff834c35acd241679ada9f08
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spelling	oai:doaj.org-article:3974c584ff834c35acd241679ada9f082021-11-30T12:07:44ZMolecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection2297-176910.3389/fvets.2021.757978https://doaj.org/article/3974c584ff834c35acd241679ada9f082021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fvets.2021.757978/fullhttps://doaj.org/toc/2297-1769Promyelocytic 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.Jingjing ZhuZhenyu ChenZhenglie DaiXiaolong ZhouHan WangXiangchen LiAyong ZhaoSongbai YangFrontiers Media S.A.articleporcinePML genealternative splicingcloneJEVVeterinary medicineSF600-1100ENFrontiers in Veterinary Science, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	porcine PML gene alternative splicing clone JEV Veterinary medicine SF600-1100
spellingShingle	porcine PML gene alternative splicing clone JEV Veterinary medicine SF600-1100 Jingjing Zhu Zhenyu Chen Zhenglie Dai Xiaolong Zhou Han Wang Xiangchen Li Ayong Zhao Songbai Yang Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection
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	article
author	Jingjing Zhu Zhenyu Chen Zhenglie Dai Xiaolong Zhou Han Wang Xiangchen Li Ayong Zhao Songbai Yang
author_facet	Jingjing Zhu Zhenyu Chen Zhenglie Dai Xiaolong Zhou Han Wang Xiangchen Li Ayong Zhao Songbai Yang
author_sort	Jingjing Zhu
title	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_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_sort	molecular cloning of alternative splicing variants of the porcine pml gene and its expression patterns during japanese encephalitis virus infection
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/3974c584ff834c35acd241679ada9f08
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Molecular Cloning of Alternative Splicing Variants of the Porcine PML Gene and Its Expression Patterns During Japanese Encephalitis Virus Infection

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