Interaction of eukaryotic proliferating cell nuclear antigen (PCNA) with the replication-associated protein (Rep) of cotton leaf curl Multan virus and pedilanthus leaf curl virus.
[en] The replication-associated (Rep) proteins of pathogenic begomoviruses, including cotton leaf curl Multan virus (CLCuMuV) and pedilanthus leaf curl virus (PeLCV), interact with the DNA replication machinery of their eukaryotic hosts. The analysis of Rep protein sequences showed that there is 13-28% sequence variation among CLCuMuV and PeLCV isolates, with phylogenetic clusters that can separated at least in part based on the country of origin of the respective viruses. To identify specific host factors involved in the virus replication cycle, we conducted yeast two-hybrid assays to detect possible interactions between the CLCuMuV and PeLCV Rep proteins and 30 protein components of the Saccharomyces cerevisiae DNA replication machinery. This showed that the proliferating cell nuclear antigen (PCNA) protein of S. cerevisiae interacts with Rep proteins from both CLCuMuV and PeLCV. We used the yeast PCNA sequence in BLAST comparisons to identify two PCNA orthologs each in Gossypium hirsutum (cotton), Arabidopsis thaliana (Arabidopsis), and Nicotiana benthamiana (tobacco). Sequence comparisons showed 38-40% identity between the yeast and plant PCNA proteins, and > 91% identity among the plant PCNA proteins, which clustered together in one phylogenetic group. The expression of the six plant PCNA proteins in the yeast two-hybrid system confirmed interactions with the CLCuMuV and PeLCV Rep proteins. Our results demonstrate that the interaction of begomovirus Rep proteins with eukaryotic PCNA proteins is strongly conserved, despite significant evolutionary variation in the protein sequences of both of the interacting partners.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Shakir, Sara ; Université de Liège - ULiège > Département GxABT > Plant Sciences ; Virology Lab, Center for Agricultural Biochemistry and Biotechnology, University of Agriculture, Faisalabad, 38000 Pakistan ; Boyce Thompson Institutute, Ithaca, NY 14853 USA
Jander, Georg; Boyce Thompson Institutute, Ithaca, NY 14853 USA
Nahid, Nazia; Department of Bioinformatics and Biotechnology, GC University, Faisalabad, Pakistan
Mubin, Muhammad; Virology Lab, Center for Agricultural Biochemistry and Biotechnology, University of Agriculture, Faisalabad, 38000 Pakistan
Younus, Ayesha; Laser Matter Interaction and Nano-Sciences Lab, Department of Physics, University of Agriculture, Faisalabad, Pakistan
Nawaz-Ul-Rehman, Muhammad Shah ; Virology Lab, Center for Agricultural Biochemistry and Biotechnology, University of Agriculture, Faisalabad, 38000 Pakistan
Language :
English
Title :
Interaction of eukaryotic proliferating cell nuclear antigen (PCNA) with the replication-associated protein (Rep) of cotton leaf curl Multan virus and pedilanthus leaf curl virus.
Publication date :
January 2021
Journal title :
3 Biotech
ISSN :
2190-572X
eISSN :
2190-5738
Publisher :
Springer Science and Business Media Deutschland GmbH, Germany
The authors are thankful to all lab members for their support in conducting research experiments. The research was funded by the International Foundation for Science (IFS grant no, C/5434-1) and the Higher Education Commission (HEC) of Pakistan (project no 1682). Sara Shakir was supported by the HEC indigenous scholarship program and the International Research Support Initiative Program (IRSIP).
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