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目的 羊口疮病毒（Orf virus, ORFV）感染可诱导皮肤角质形成细胞（Keratinocyte cell, KC）发生不全性角化过度（Parakeratotic hyperkeratosis, PHK），其中的分子机制尚不清楚。CDC42(Cell division cycle 42)是Rho GTPase家族成员之一，能够协同磷脂酶D(Phospholipase D, PLD)，发挥重要的细胞分化调控功能。本研究将验证病毒晚期囊膜蛋白B2L(Viral PLD, vPLD)与细胞CDC42分子是否存在互作关系，为深入研究B2L促进KC分化的分子机制奠定基础。方法 基于生物信息学预测B2L与CDC42分子空间的互作形式；进而利用RT-PCR、Western blot和免疫共沉淀技术（Co-immunoprecipitation, Co-IP）等方法验证二者之间的作用及其依存关系。结果CDC42与B2L分子能够形成空间稳定的复合物，并且二者在mRNA和蛋白水平均存在剂量依赖性。结论 本研究揭示了ORFV晚期蛋白B2L(vPLD)能够结合细胞CDC42分子，并影响其表达水平。这为进一步探索ORFV诱导KC分化及其PHK损伤的分子机制奠定了基础。
Abstract：Objective Orf virus(ORFV) infection induces parakeratotic hyperkeratosis(PHK) in skin keratinocytes(KC), yet the underlying molecular mechanisms remain unclear. Cell division cycle 42(CDC42), a member of the Rho GTPase family, cooperates with phospholipase D(PLD) to play an essential role in regulating cell differentiation. This study aims to determine whether an interaction exists between the ORFV late envelope protein B2L(viral phospholipase D, vPLD) and the cellular CDC42, thereby providing a basis for elucidating the molecular mechanisms by which B2L promotes KC differentiation. Methods Bioinformatics analyses were first performed to predict the potential spatial interaction between B2L and CDC42. Subsequently, reverse transcription – polymerase chain reaction(RT-PCR), Western blot, and coimmunoprecipitation(Co-IP) assays were employed to verify their interaction and to assess the dependence of their expression. Results B2L and CDC42 formed a stable molecular complex. Moreover, their expression exhibited dose-dependent changes at both the mRNA and protein levels.Conclusion This study demonstrates that the ORFV late protein B2L(vPLD) interacts with cellular CDC42 and modulates its expression. These findings provide a molecular basis for further investigation into ORFV-induced keratinocyte differentiation and the development of PHK.
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基本信息:
中图分类号:S852.65
引用信息:
[1]黄欣,艾书晴,何鑫,等.羊口疮病毒B2L蛋白诱导细胞CDC42分子的表达[J].病毒学报().
基金信息:
国家自然科学基金面上项目(项目号:31172353),题目:羊传染性脓疱病毒保护性抗原表位筛选及表位作图研究;国家自然科学基金面上项目(项目号:32272991),题目:长链非编码RNA MSTRG.22610.1促进BHV-1在MDBK细胞中增殖的分子机制; 黑龙江省教育厅课题(项目号:11551322),题目:口蹄疫病毒Ca2+信号通道结构的研究
2026-02-24
2026-02-24
2026-02-24