Adeno-associated Virus 9 Structural Rearrangements Induced by Endosomal Trafficking pH and Glycan Attachment

内吞作用 内吞循环 病毒进入 糖蛋白 高尔基体 病毒 脂质双层融合 病毒包膜
作者
Judit J Pénzes,Paul R. Chipman,Nilakshee Bhattacharya,Allison Zeher,Rick Huang,Robert McKenna,Mavis Agbandje-McKenna
出处
期刊:Journal of Virology [American Society for Microbiology]
卷期号:95 (19) 被引量:2
标识
DOI:10.1128/jvi.00843-21
摘要

Adeno-associated viruses (AAVs) are small nonenveloped single-stranded DNA (ssDNA) viruses that are currently being developed as gene therapy biologics. After cell entry, AAVs traffic to the nucleus using the endo-lysosomal pathway. The subsequent decrease in pH triggers conformational changes to the capsid that enable the externalization of the capsid protein (VP) N termini, including the unique domain of the minor capsid protein VP1 (VP1u), which permits the phospholipase activity required for the capsid lysosomal egress. Here, we report the AAV9 capsid structure, determined at the endosomal pHs (7.4, 6.0, 5.5, and 4.0), and terminal galactose-bound AAV9 capsids at pHs 7.4 and 5.5 using cryo-electron microscopy and three-dimensional image reconstruction. Taken together, these studies provide insight into AAV9 capsid conformational changes at the 5-fold pore during endosomal trafficking, in both the presence and absence of its cellular glycan receptor. We visualized, for the first time, that acidification induces the externalization of the VP3 and possibly VP2 N termini, presumably in prelude to the externalization of VP1u at pH 4.0, which is essential for lysosomal membrane disruption. In addition, the structural study of AAV9-galactose interactions demonstrates that AAV9 remains attached to its glycan receptor at the late endosome pH 5.5. This interaction significantly alters the conformational stability of the variable region I of the VPs, as well as the dynamics associated with VP N terminus externalization. IMPORTANCE There are 13 distinct Adeno-associated virus (AAV) serotypes that are structurally homologous and whose capsid proteins (VP1 to -3) are similar in amino acid sequence. However, AAV9 is one of the most commonly studied and is used as a gene therapy vector. This is partly because AAV9 is capable of crossing the blood-brain barrier and readily transduces a wide array of tissues, including the central nervous system. In this study, we provide AAV9 capsid structural insight during intracellular trafficking. Although the AAV capsid has been shown to externalize the N termini of its VPs, to enzymatically disrupt the lysosome membrane at low pH, there was no structural evidence to confirm this. By utilizing AAV9 as our model, we provide the first structural evidence that the externalization process occurs at the protein interface at the icosahedral 5-fold symmetry axis and can be triggered by lowering the pH.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
HHHAN发布了新的文献求助10
3秒前
胡胡完成签到 ,获得积分10
4秒前
火星上小土豆完成签到 ,获得积分10
14秒前
杰尼龟的鱼完成签到 ,获得积分10
18秒前
安然完成签到 ,获得积分10
19秒前
张希伦完成签到 ,获得积分10
19秒前
任性翠安完成签到 ,获得积分10
23秒前
dong完成签到 ,获得积分10
25秒前
神说完成签到,获得积分0
27秒前
量子星尘发布了新的文献求助10
33秒前
Aimee完成签到 ,获得积分10
33秒前
徐彬荣完成签到,获得积分10
34秒前
研友_8yN60L完成签到,获得积分10
35秒前
搜集达人应助科研通管家采纳,获得10
35秒前
光亮的自行车完成签到 ,获得积分10
35秒前
李东东完成签到 ,获得积分10
50秒前
王多肉完成签到,获得积分10
54秒前
Iiiilr完成签到 ,获得积分10
55秒前
杨幂完成签到,获得积分10
56秒前
1分钟前
hellokitty完成签到,获得积分10
1分钟前
1分钟前
小四发布了新的文献求助10
1分钟前
1分钟前
西瓜完成签到 ,获得积分10
1分钟前
包容的忆灵完成签到 ,获得积分10
1分钟前
高兴尔冬发布了新的文献求助10
1分钟前
xiang完成签到 ,获得积分0
1分钟前
小四完成签到,获得积分10
1分钟前
FashionBoy应助slayers采纳,获得30
1分钟前
量子星尘发布了新的文献求助10
1分钟前
黑眼圈完成签到 ,获得积分10
1分钟前
jia完成签到 ,获得积分10
1分钟前
如履平川完成签到 ,获得积分10
1分钟前
科目三应助忧伤的步美采纳,获得10
1分钟前
大椒完成签到 ,获得积分10
1分钟前
1分钟前
1分钟前
wisdom完成签到,获得积分10
1分钟前
slayers发布了新的文献求助30
1分钟前
高分求助中
【提示信息,请勿应助】关于scihub 10000
Les Mantodea de Guyane: Insecta, Polyneoptera [The Mantids of French Guiana] 3000
徐淮辽南地区新元古代叠层石及生物地层 3000
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
Handbook of Industrial Diamonds.Vol2 1100
Global Eyelash Assessment scale (GEA) 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 550
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 4038039
求助须知:如何正确求助?哪些是违规求助? 3575756
关于积分的说明 11373782
捐赠科研通 3305574
什么是DOI,文献DOI怎么找? 1819239
邀请新用户注册赠送积分活动 892655
科研通“疑难数据库(出版商)”最低求助积分说明 815022