A Novel Substrate of Src Tyrosine Kinases

描述(由申请人提供)：Fyn酪氨酸激酶，成员 Src家族，促进角质形成细胞分化。角质形成细胞来源 FYN缺乏的小鼠表现出鳞片形成受损，并显著减少 分化标志物如微丝蛋白和转谷氨酰胺酶的表达。 此外，Fyn缺乏的角质形成细胞表现出酪氨酸磷酸化水平降低。 连环蛋白是正常的细胞间黏附所必需的。理解 Fyn如何诱导角质形成细胞分化据说是 在分子水平上了解皮肤病。他们假设芬恩 可能通过与其他分子相互作用和磷酸化来发挥作用。 为了解决这个问题，他们对一只小鼠进行了酵母双杂交筛选 以FYN为诱饵的角质形成细胞库。这一幕产生了一部小说 被Fyn磷酸化并与其结合的假定的接头分子； 他们将这种分子命名为srCasm：src激活和信号分子。他们 建议用生化方法表征Fyn和srCasm之间的相互作用 并阐明Fyn磷酸化和Fyn磷酸化所需的srCasm残基和 协会。他们还将研究SRCasm如何与其他信号相关联 SrCasm分子及其在上皮和人中的表达 皮肤病。随着腺病毒产生各种形式的Fyn和Src， 他们将评估sRCasm在信号转导途径中的作用 初级角质形成细胞中的这些激酶。最后，由于他们的加入， 原位研究，他们将针对部分表皮表达srCasm 和毛囊中它不表达的地方，并将检查其影响 异常的srCasm表达。 以下是临床科学家发展导师奖的申请 设想促进候选人从博士后研究员过渡到 一名独立调查员。候选人将把85%的精力投入到研究中 设计用来表征一种新的Src家族激酶底物在 角质形成细胞信号转导。这项研究将在该部门进行 有支持性环境的大型教学医院的皮肤科 以及致力于内科科学家的专业发展。这个 申请者的其余工作，即15%，将涉及诊断皮肤活检。

DESCRIPTION (provided by the applicant): The Fyn tyrosine kinase, a member of the Src family, promotes keratinocyte differentiation. Keratinocytes from Fyn-deficient mice exhibit impaired squame formation, and markedly decreased expression of differentiation markers such as filaggrin and transglutaminase. Also, Fyn-deficient keratinocytes show decreased tyrosine phosphorylation of catenins which are necessary for proper intercellular adhesion. Understanding how Fyn induces keratinocyte differentiation is said to be fundamental for understanding cutaneous disease at a molecular level. They hypothesize that Fyn may exert its effects by interacting with and phosphorylating other molecules. To address this question, they performed a yeast two-hybrid screen of a murine keratinocyte library using Fyn as the bait. This screen yielded a novel putative adaptor molecule which is phosphorylated by and associates with Fyn; they term this molecule Srcasm: Src activating and signaling molecule. They propose to biochemically characterize the interaction between Fyn and Srcasm and elucidate what residues in Srcasm are required for Fyn phosphorylation and association. They will also examine how Srcasm associates with other signaling molecules and characterize the expression of Srcasm in epithelial and human cutaneous diseases. With adenoviruses producing various forms of Fyn and Src, they will evaluate the role of Srcasm in the signal-transduction pathways of these kinases within primary keratinocytes. Lastly, as a result of their in situ studies, they will target expression of Srcasm to parts of the epidermis and hair follicle where it is not expressed and will examine the effects of aberrant Srcasm expression. This application for a Mentored Clinical Scientist Development Award is envisaged to promote the candidate's transition from a postdoctoral fellow to an independent investigator. The candidate will devote 85% effort to research designed to characterize the role of a novel Src family kinase substrate in keratinocyte signal transduction. The research will be done in the Department of Dermatology of a major teaching hospital which has a supportive environment and a commitment to the professional development of physician scientists. The rest of the applicant's effort, 15%, will involve diagnosing skin biopsies.