Non Fiber Cell Functions for the Beaded Filament Proteins

珠状丝蛋白的非纤维细胞功能

• 批准号: 8747592
• 负责人: PAUL Gillespie FITZGERALD
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8747592
• 关键词: Affinity Chromatography; Age; Animals; Anterior; Antibodies; Ants; Appearance; Attention; Biological; Biology; Biopsy; Cell Cycle; Cell division; Cell physiology; Cells; Cellular biology; Complementary DNA; Crystalline Lens; Crystallins; Cytokinesis; Cytoskeletal Filaments; DNA; DNA biosynthesis; Data; Development; Epithelial; Epithelial Cells; Epithelium; Filament; Future; Genetic Engineering; Human; In Situ Hybridization; Intermediate Filaments; Knock-out; Knowledge; Label; Lens Fiber; Link; Literature; Malignant Neoplasms; Mass Spectrum Analysis; Mitosis; Mitotic Activity; Molecular; Molecular Chaperones; Mus; Organelles; Pathologic; Population; Process; Proteins; Relative (related person); Reporting; Resources; S-Phase Fraction; Structure; Surgical sutures; System; Thymidine; Time; Tissues; Transcend; Translating; Translations; Ubiquitin; Vimentin; Work; analog; base; daughter cell; experience; fiber cell; filensin; knockout animal; lens; lens protein; novel; nuclear division; protein expression; protein function; public health relevance; tool; tumor

Abstract Beaded Filaments (BFs) are a structurally and biochemically filamentous cytoskeletal structure found in lens fiber cells. In 40 years, neither the BF nor the two proteins required for assembly of the BF (BF Proteins CP49 and filensin), have been demonstrated in cells other than differentiated lens fiber cells. In Preliminary Data, we show that BF proteins are expressed in lens epithelium, but also in a subset of biopsied human cancers. Moreover, BF protein expression in lens epithelium is not in a filamentous structure, but instead in a large vermiform structure, which is only expressed after 5 weeks of age. As cells near the bow region where differentiation is triggered, the structure disappears. These data: 1) are the first to report (to our knowledge) the expression of BF proteins outside of the fiber cell, and outside of the lens 2) suggest that the lens epithelium may assemble a novel organelle/inclusion, and 3) that developmental switches are still being flipped in the lens epithelium at 5 weeks of age. Thus, these observations suggest a previously unknown function for the BF Proteins, both in the lens, and in non-lens tissue, and in both normal and pathologic biology. We have no idea what this function is, and invoke this R21 to begin the process of establishing this/these function(s). Our lab has worked on Beaded Filaments and Intermediate Filaments for the past 25 years. In doing so we have accumulated a combination of BF/IF tools, resources, and experience in lens biology, which uniquely situates us to make progress in revealing this "non-canonical" BF protein function. Because of the availability of these tools (CP49 knockout, filensin knockout, and vimentin-knockout animals, a range of antibodies to BF proteins, vimentin, and several other lens proteins, cDNA constructs, etc) and because of the limited availability of rare human tumor material, we focus these initial efforts on lens. There are, of course, a vast number of questions that can be asked about a novel structure, but the R21 limitations constrain the scope of inquiry. For that reason we are proposing questions at three levels, the molecular, cellular and tissue levels, to cast the widest net possible. The questions asked are guided by our knowledge of lens, intermediate filaments, beaded filaments, and the literature on cellular organelles and inclusions. Just as our previous work started with a fiber cell-specific cytoskeletal structure (the BF) and grew into a general attack on the structure of Intermediate Filaments, we hope that these studies will transcend from lens epithelial-specific to the broader field of cell biology.

摘要 珠状纤维（BF）是一种结构和生化丝状细胞骨架结构，发现于 透镜纤维细胞。40年来，无论是BF还是组装BF所需的两种蛋白质（BF Proteins CP 49和filensin），已经在除分化的透镜纤维细胞之外的细胞中得到证实。初步 数据，我们表明BF蛋白表达在透镜上皮细胞，但也在一个子集的活检人类晶状体上皮细胞， 癌的此外，BF蛋白在透镜上皮中的表达不是以丝状结构，而是以丝状结构表达。 大的蠕虫状结构，仅在5周龄后表达。作为靠近弓形区域的细胞， 分化被触发，结构消失。这些数据：1）是第一次报告（据我们所知） BF蛋白在纤维细胞外和透镜外的表达2）表明透镜上皮细胞 可能组装一个新的细胞器/内含物，3）发育开关仍在透镜中翻转 5周龄时的上皮细胞。因此，这些观察结果表明了BF的一个以前未知的功能 蛋白质，包括透镜和非透镜组织中的蛋白质，以及正常和病理生物学中的蛋白质。我们不知道 此功能是什么，并调用此R21以开始建立此/这些功能的过程。 我们的实验室在过去的25年里一直致力于珠状长丝和中间长丝的研究。做 因此，我们在透镜生物学方面积累了BF/IF工具、资源和经验， 独特地使我们在揭示这种“非经典”BF蛋白功能方面取得进展。因为 这些工具的可用性（CP 49敲除、filensin敲除和vimentin敲除动物，一系列 BF蛋白、波形蛋白和几种其它透镜蛋白、cDNA构建体等的抗体），并且由于 由于罕见的人类肿瘤材料的有限可用性，我们将这些最初的努力集中在透镜上。 当然，对于一个新颖的结构，可以提出大量的问题，但是， R21的局限性限制了调查的范围。因此，我们提出了三个层次的问题， 分子、细胞和组织水平，尽可能广泛地撒网。所提出的问题是由我们的 知识的透镜，中间丝，串珠丝，和文献的细胞器和 内含物。 就像我们以前的工作从纤维细胞特异性的细胞骨架结构（BF）开始， 对中间纤维结构的一般性研究，希望这些研究能超越透镜的范畴 上皮特异性的细胞生物学更广泛的领域。