Effects of transcription units and a specific hexanucleotide motif of porcine circovirus-like virus P1 on viral DNA synthesis, viral protein expression, and progeny virus production.
15 affiliations
- 1College of Animal Science, Provincial Key Laboratory of Tibet Plateau Animal Epidemic Disease Research, Tibet Agricultural and Animal Husbandry University, Linzhi, 860000, China.
- 2Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.
- 3Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture and Rural Affairs, Nanjing, 210014, China.
- 4Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infections Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
- 5Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, 210014, China.
- 6Huai'an Animal Disease Prevention and Control Center, Huai'an, 223001, China.
- 7College of Animal Science, Provincial Key Laboratory of Tibet Plateau Animal Epidemic Disease Research, Tibet Agricultural and Animal Husbandry University, Linzhi, 860000, China. [email protected].
- 8Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China. [email protected].
- 9Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture and Rural Affairs, Nanjing, 210014, China. [email protected].
- 10Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infections Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China. [email protected].
- 11Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, 210014, China. [email protected].
- 12Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China. [email protected].
- 13Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture and Rural Affairs, Nanjing, 210014, China. [email protected].
- 14Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infections Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China. [email protected].
- 15Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing, 210014, China. [email protected].
Abstract
Background
Porcine circovirus-like virus P1 has a broad host range and is known to cause post-weaning multisystemic wasting syndrome in pigs, as well as infect and propagate in cattle, goats, dogs, cats, yaks, and rabbits. To investigate the functional importance of its eight open reading frames (ORFs) and the conserved hexanucleotide motif (GGATCC), we generated a panel of site-directed mutants based on the P1 infectious clone rpSK-2P1. These mutants were transfected into PK-15 cells and serially passaged four times. Viral replication capacity and genetic stability were assessed using polymerase chain reaction (PCR), quantitative PCR, and Sanger sequencing.Results
All the mutant viruses retained the ability to replicate their genomes and transcribe viral RNA. However, sequencing of viral genomes recovered at the fourth passage revealed selective reversion at multiple mutated sites, indicating a strong replicative advantage conferred by wild-type sequences during sustained propagation. Notably, mutant Y1, which harbored a mutation in ORF1 that encoded the capsid protein (Cap), initially exhibited reduced replication but rapidly reverted upon passaging, restoring Cap expression and fitness.Conclusions
These findings demonstrate that although these ORFs and nucleotide sites do not abolish P1 replication, their native sequences play pivotal roles in modulating replication efficiency and maintaining genetic stability, thereby providing key insights into the molecular mechanisms governing P1 replication. Thus, P1 tolerates limited genetic change but is under intense pressure to maintain its wild-type sequence, offering critical insight into the replication strategy and evolutionary constraints of this compact, broad-host-range DNA virus.
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BibTeX
@article{song2026effects,
title = {{Effects of transcription units and a specific hexanucleotide motif of porcine circovirus-like virus P1 on viral DNA synthesis, viral protein expression, and progeny virus production.}},
author = {Jingze Song and Zixuan Li and Na Li and Jianping Xie and Hengzhi Zhu and Hongqiao Lai and Zenghao Xiang and Yanan Zhong and Hao Sun and Kongwang He and Sizhu Suolang and Libin Wen and Qi Xiao},
journal = {BMC veterinary research},
year = {2026},
month = jul,
doi = {10.1186/s12917-026-05740-0},
url = {https://europepmc.org/article/MED/42928309},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Effects of transcription units and a specific hexanucleotide motif of porcine circovirus-like virus P1 on viral DNA synthesis, viral protein expression, and progeny virus production.
AU - Jingze Song
AU - Zixuan Li
AU - Na Li
AU - Jianping Xie
AU - Hengzhi Zhu
AU - Hongqiao Lai
AU - Zenghao Xiang
AU - Yanan Zhong
AU - Hao Sun
AU - Kongwang He
AU - Sizhu Suolang
AU - Libin Wen
AU - Qi Xiao
PY - 2026
DA - 2026/07/28
JO - BMC veterinary research
DO - 10.1186/s12917-026-05740-0
UR - https://europepmc.org/article/MED/42928309
AB - Background Porcine circovirus-like virus P1 has a broad host range and is known to cause post-weaning multisystemic wasting syndrome in pigs, as well as infect and propagate in cattle, goats, dogs, cats, yaks, and rabbits. To investigate the functional importance of its eight open reading frames (ORFs) and the conserved hexanucleotide motif (GGATCC), we generated a panel of site-directed mutants based on the P1 infectious clone rpSK-2P1. These mutants were transfected into PK-15 cells and serially passaged four times. Viral replication capacity and genetic stability were assessed using polymerase chain reaction (PCR), quantitative PCR, and Sanger sequencing.Results All the mutant viruses retained the ability to replicate their genomes and transcribe viral RNA. However, sequencing of viral genomes recovered at the fourth passage revealed selective reversion at multiple mutated sites, indicating a strong replicative advantage conferred by wild-type sequences during sustained propagation. Notably, mutant Y1, which harbored a mutation in ORF1 that encoded the capsid protein (Cap), initially exhibited reduced replication but rapidly reverted upon passaging, restoring Cap expression and fitness.Conclusions These findings demonstrate that although these ORFs and nucleotide sites do not abolish P1 replication, their native sequences play pivotal roles in modulating replication efficiency and maintaining genetic stability, thereby providing key insights into the molecular mechanisms governing P1 replication. Thus, P1 tolerates limited genetic change but is under intense pressure to maintain its wild-type sequence, offering critical insight into the replication strategy and evolutionary constraints of this compact, broad-host-range DNA virus.
SN - 1746-6148
ER -