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[The N1-18 terminus of Marek's disease virus VP22 is essential for protein transduction]
Hongjun Chen1, Aijian Qin, Cuiping Song
1Jiangsu Animal Preventive Medicine Key Lab, Yangzhou University, China 225009.)
Abstract:
We previously showed some differences in Marek's disease virus (MDV) VP22 gene between virulent and avirulent strains, in the deletion from 201aa to 206aa, namely 201TKSERT206. In this study, VP22 genes were amplified from strains: CVI988/Rispens and GA. And then the fragments were subcloned into pcDNA3.1/zeo(+), respectively, which were co-expressed with an enhancer green fluorescent protein (EGFP) after transfection into COS-1 cells. As with both human herpesvirus 1 and bovine herpesvirus 1 VP22-EGFP fusion proteins, the subcellular localization of the three MDV EGFP-VP22 products revealed few differences, which bind to microtubules and nucleus membrane, and then to heterochromatin. In addition, VP22s also bind to centrosomes and inter-membrane. During mitosis, EGFP-VP22s bind to sister chromatids, but dissociates from the centrosomes and the microtubules of the mitotic spindle. In truncated fragments' transfection experiments, stained with the specific monoclonal antibody against VP22, it concluded that the full length of VP22 was required for protein transduction, and N1-18aa was essential to VP22 translocating from cytoplasm to nucleus as a potential nucleus localization site in the absence of other viral factors in MDV-1.
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