Early events of in vitro KSHV infection

体外 KSHV 感染的早期事件

基本信息

批准号：
8446318
负责人：
Bala Chandran
金额：
$ 30.14万
依托单位：
ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8446318
关键词：
Acetylation Actins Address Animal Viruses B-Lymphocytes Binding Bulla Capsid Cell Nucleus Cell Surface Receptors Cell surface Cells Clathrin Complex Dermal Disease Dynein ATPase Endocytosis Endothelial Cells Ephrin-A2 Event Fibroblasts Gene Expression Germ Cells Goals Guanosine Triphosphate Phosphohydrolases Heparitin Sulfate Herpesviridae Herpesviridae Infections Human Human Herpesvirus 8 In Vitro Infection Integrins Kaposi Sarcoma Link MAPK3 gene Malignant Neoplasms Mediating Membrane Microdomains Microtubules Mono-S Motor Multicentric Angiofollicular Lymphoid Hyperplasia NF-kappa B Nonmuscle Myosin Type IIA Nuclear PTK2 gene Pathway interactions Play Protein Tyrosine Kinase Proteins Recruitment Activity Research Role Signal Pathway Signal Transduction Signaling Molecule Sorting - Cell Movement Staging Testing Ubiquitination Umbilical vein Viral Viral Genes Virus Virus Activation bindin cell motility ezrin monocyte novel primary effusion lymphoma receptor rho GTP-Binding Proteins viral DNA

项目摘要

DESCRIPTION (provided by applicant): There is a fundamental gap in our understanding of how animal virus interactions with cell surface receptors facilitate a productive infection. Bindin of viruses, such as herpesviruses, to the cell surface can occur at 40C in an energy independent manner. In contrast, entry into cells and subsequent transfer of capsids to the vicinity of the nucleus are an active, energy dependent phenomenon, and thus must require host-cell signaling pathways. Our central hypothesis is that KSHV has evolved to utilize its interactions with receptors to manipulate the host's pre-existing signal cascades to mediate entry and infection of target cells. Our ongoing studies support this hypothesis. We have shown that during infection of endothelial (HMVEC-d) cells, the initial attachment of KSHV occurs via heparan sulfate (HS) followed by a temporal interaction with integrins (?3?1,??V??3, and ?V??5) and xCT molecules. KSHV binding resulted in the activation of FAK, Src, PI3-K, Rho-GTPases, Dia-2, Ezrin, PKC-?, ERK1/2 and NF-?B signal molecules. These molecules play roles in KSHV entry (FAK, Src, PI3-K, Rho-GTPases), acetylation of microtubules (MT) (RhoA-GTPase) that facilitates the transport of KSHV capsid toward the nucleus via dynein motors, and viral gene expression (ERK1/2 and NF-?B). Although lipid raft (LR) disruption resulted in increased Src activation and virus entry, we observed the inhibition of viral gene expression, PI3K, RhoA, Dia-2 and NF?B activation, MT acetylation and nuclear delivery of viral DNA which suggested that LRs play roles in entry in cells and modulates selected signal molecules. We have also demonstrated that c-Cbl play a role in KSHV macropinocytosis. To further test our hypothesis, we have formulated three major interlinked, focused specific aims which will decipher the mechanisms by which c-Cbl facilitate KSHV entry, the role of major adaptor molecules in KSHV infection and decipher how c-Cbl dictates the fate of macropinosomes containing KSHV. These studies are significant since such comprehensive understanding of early events of KSHV infection will provide novel targets to block the initiation of target cell infection by KSHV and te associated diseases.

描述（由申请人提供）：我们对动物病毒与细胞表面受体的相互作用如何促进生产感染的理解存在基本差距。病毒（例如疱疹病毒）与细胞表面的绑定可以以独立的方式出现在40℃。相比之下，进入细胞并随后将衣壳传递到核附近的转移是一种活跃的，依赖能量的现象，因此必须需要宿主 - 细胞信号传导途径。我们的中心假设是，KSHV已经发展为利用其与受体的相互作用，以操纵宿主先前存在的信号级联反应，以介导目标细胞的进入和感染。我们正在进行的研究支持这一假设。我们已经表明，在感染内皮（HMVEC-D）细胞期间，KSHV的初始附着是通过硫酸乙酰肝素（HS）发生的，然后与整合素（？3？1，?? V ?? 3和？v ?? 5）和XCT分子发生时间相互作用。 KSHV结合导致FAK，SRC，PI3-K，Rho-GTPases，DIA-2，Ezrin，Pkc-？，erk1/2和NF-？B信号分子的激活。这些分子在KSHV进入（FAK，SRC，PI3-K，Rho-GTPases），微管（MT）（Rhoa-GTPase）的乙酰化中扮演角色，可促进KSHV通过Dynein Motors通过Dynein Motors和Veral Gene表达（ERK1/2/2和NF）的KSHV capsid朝向核的运输。尽管脂质筏（LR）破坏导致SRC激活和病毒的进入增加，但我们观察到病毒基因表达，PI3K，RHOA，DIA-2和NF？B激活，MT乙酰化，MT乙酰化和核传递病毒DNA的抑制作用，该病毒DNA表明LRS在细胞和调节中所选择的信号分子中发挥了发挥作用。我们还证明了C-CBL在KSHV大型细胞增多症中起作用。为了进一步检验我们的假设，我们制定了三个主要相互关注的特定目的，这将破译C-CBL促进KSHV进入的机制，即KSHV分子在KSHV感染中的作用，并破译C-CBL如何决定含有KSHV的大分子症的效果。这些研究很重要，因为对KSHV感染的早期事件的这种全面理解将提供新的靶标，以阻止KSHV和TE相关疾病引发靶细胞感染。