Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses

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February 2005

Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses[J]. Virologica Sinica, 2005, 20(1): 20-23.

Citation: . Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses .VIROLOGICA SINICA, 2005, 20(1) : 20-23.

人多瘤病毒BK株VP1的C端阳电荷残基对病毒样颗粒形成的影响

1.
1. 广东省疾病预防控制中心广东广州 2.日本国立感染症研究所日本东京

摘要

VP1是人多瘤病毒BK株的主要结构蛋白,使用重组杆状病毒表达系统在体外表达VP1 可以形成病毒样颗粒(VLP)。为了探讨VP1的C末端阳电荷残基R 281, R 285, K 288, R 290, R 292, K 293, R 294,和K297 对VLP形成和其结合DNA的影响,我们分别改变将阳电荷残基变成丙氨酸,然后表达VP1 蛋白。结果发现用丙氨酸替代K 288,R 290,R 292,K 293,R 294后仍能形成VLP, 但与野毒株相比,在VLP分泌以及衣壳蛋白与细胞DNA的结合方面有差异。有趣的是,R 281被丙氨酸取代后仅在细胞中形成少量的VLP,而R 285 被丙氨酸取代后不能形成VLP。该研究证实阳电荷氨基酸残基R 281 和R 285 是形成VLP所必须的,K 288、R 290、R 292、K 293、R 294和K 297则影响VLP和DNA的结合。
- 人多瘤病毒BK株
- , 病毒样颗粒
- , 杆状病毒表达
- , 精氨酸

Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses

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Abstract

VP1 is the major structural protein of the human polyomavirus BK and can be used for the generation of virus-like particles (VLP) in vitro by using recombinant baculoviruses. To determine the role of positively charged residues R-281, R-285, K-288, R-290, R-292, K-293, R-294, and K297 at the C terminal of VP1 in VLP formation and DNA binding, we separately changed the residue to alanine, and expressed VP1 protein. The results showed that substitution of K-288, R-290, R-292, K-293, R-294, and K297 with alaninecan still form VLP, but compared with wild type there were some differences in releasing VLP into culture medium and binding between major capsid proteins and cellular DNA . Interestingly,after substituting R-281 and R-285 with alanine, a few VLP in cell but no VLP were detected respectively. This study provides evidence that positively charged residues R-281 and R-285 are necessary to form VLP and the others can influence the binding between major capsid proteins and cellular DNA.
- Polyomavirus BK strain
- , Virus-like particle
- , Arginine residue
- , Baculovirus expression

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses

^1* ,
² ,
²

Abstract: VP1 is the major structural protein of the human polyomavirus BK and can be used for the generation of virus-like particles (VLP) in vitro by using recombinant baculoviruses. To determine the role of positively charged residues R-281, R-285, K-288, R-290, R-292, K-293, R-294, and K297 at the C terminal of VP1 in VLP formation and DNA binding, we separately changed the residue to alanine, and expressed VP1 protein. The results showed that substitution of K-288, R-290, R-292, K-293, R-294, and K297 with alaninecan still form VLP, but compared with wild type there were some differences in releasing VLP into culture medium and binding between major capsid proteins and cellular DNA . Interestingly,after substituting R-281 and R-285 with alanine, a few VLP in cell but no VLP were detected respectively. This study provides evidence that positively charged residues R-281 and R-285 are necessary to form VLP and the others can influence the binding between major capsid proteins and cellular DNA.