Herpes simplex virus 1 protein pUL21 alters ceramide metabolism by activating the interorganelle transport protein CERT.
Herpes simplex virus 1 protein pUL21 alters ceramide metabolism by activating the interorganelle transport protein CERT.
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DOI：
10.1016/j.jbc.2022.102589
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发表时间：
2022-11
影响因子：
4.8
通讯作者：
Graham, Stephen C.
中科院分区：
文献类型：
--
作者：
Benedyk, Tomasz H.;Connor, Viv;Caroe, Eve R.;Shamin, Maria;Svergun, Dmitri I.;Deane, Janet E.;Jeffries, Cy M.;Crump, Colin M.;Graham, Stephen C.
单纯疱疹病毒（HSV）-1在感染过程中显著改变细胞膜的结构和蛋白质组成，但其对膜脂质组成的影响尚不清楚。HSV-1 pUL 21是一种病毒编码的蛋白磷酸酶接头，可促进多种细胞和病毒蛋白的去磷酸化，包括细胞神经酰胺（Cer）转运蛋白CERT。CERT介导非囊泡Cer从内质网转运到trans-Golgi网络，随后Cer转化为鞘磷脂（SM）和其他鞘脂，在细胞增殖、信号传导和膜运输中发挥重要作用。在这里，我们使用点击化学分析鞘脂代谢的动力学，表明pUL 21介导的去磷酸化激活CERT并加速Cer-to-SM转化。纯化的pUL 21和全长CERT相互作用亚微摩尔的亲和力，我们解决了溶液结构的pUL 21 C-末端结构域的复杂的CERT Pleckstrin同源性和类固醇生成急性调节相关的脂质转移结构域使用小角X-射线散射。我们确定了一个单一的氨基酸突变的pUL 21的表面上，破坏CERT结合在体外和培养的细胞。该残基在单纯病毒属中高度保守。此外，我们确定了一个pUL 21残基的结合HSV-1 pUL 16。鞘脂分析表明，在HSV-1感染的情况下，Cer-to-SM转化严重减少，当感染编码缺乏激活CERT能力的突变形式pUL 21的病毒时，这种缺陷会加剧。然而，当pUL 21介导的CERT去磷酸化被消除时，培养的角质形成细胞或上皮细胞中的病毒复制和传播并未显着改变。总的来说，我们证明了HSV-1通过特定的蛋白质-蛋白质相互作用改变鞘脂代谢。
Herpes simplex virus (HSV)-1 dramatically alters the architecture and protein composition of cellular membranes during infection, but its effects upon membrane lipid composition remain unclear. HSV-1 pUL21 is a virus-encoded protein phosphatase adaptor that promotes dephosphorylation of multiple cellular and virus proteins, including the cellular ceramide (Cer) transport protein CERT. CERT mediates nonvesicular Cer transport from the endoplasmic reticulum to the trans-Golgi network, whereupon Cer is converted to sphingomyelin (SM) and other sphingolipids that play important roles in cellular proliferation, signaling, and membrane trafficking. Here, we use click chemistry to profile the kinetics of sphingolipid metabolism, showing that pUL21-mediated dephosphorylation activates CERT and accelerates Cer-to-SM conversion. Purified pUL21 and full-length CERT interact with submicromolar affinity, and we solve the solution structure of the pUL21 C-terminal domain in complex with the CERT Pleckstrin homology and steroidogenic acute regulatory–related lipid transfer domains using small-angle X-ray scattering. We identify a single amino acid mutation on the surface of pUL21 that disrupts CERT binding in vitro and in cultured cells. This residue is highly conserved across the genus Simplexvirus. In addition, we identify a pUL21 residue essential for binding to HSV-1 pUL16. Sphingolipid profiling demonstrates that Cer-to-SM conversion is severely diminished in the context of HSV-1 infection, a defect that is compounded when infecting with a virus encoding the mutated form of pUL21 that lacks the ability to activate CERT. However, virus replication and spread in cultured keratinocytes or epithelial cells is not significantly altered when pUL21-mediated CERT dephosphorylation is abolished. Collectively, we demonstrate that HSV-1 modifies sphingolipid metabolism via specific protein–protein interactions.
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影响因子：
11.3
作者：
Bigalke JM;Heldwein EE
通讯作者：
Heldwein EE
影响因子：
11.4
作者：
Bigalke, Janna M.;Heldwein, Ekaterina E.
通讯作者：
Heldwein, Ekaterina E.
影响因子：
4.4
作者：
Bell, Christina;Desjardins, Michel;Radtke, Kerstin
通讯作者：
Radtke, Kerstin
影响因子：
6.1
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Blanchet CE;Spilotros A;Schwemmer F;Graewert MA;Kikhney A;Jeffries CM;Franke D;Mark D;Zengerle R;Cipriani F;Fiedler S;Roessle M;Svergun DI
通讯作者：
Svergun DI
影响因子：
10.5
作者：
Chuluunbaatar, Uyanga;Roller, Richard;Mohr, Ian
通讯作者：
Mohr, Ian