BIOGENESIS OF THE RED CELL MEMBRANE AND CYTOSKELETON

红细胞膜和细胞骨架的生物发生

• 批准号: 3087622
• 负责人: HAGOP YOUSSOUFIAN
• 金额: $ 9.34万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-10-01 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3087622
• 关键词: actins; ankyrins; cell differentiation; cytoskeleton; erythrocyte membrane; erythropoiesis; gelsolin; laboratory mouse; membrane biogenesis; polymerization; protein signal sequence; protein transport; spectrin; tissue /cell culture; transfection

Mechanisms of protein sorting are of fundamental importance in differentiation, and mammalian erythropoiesis provides a valuable model to study this phenomenon. The erythroid proteins spectrin and band 3, for example, remain with the mature reticulocyte, while most surface proteins segregate with the extruded nucleus. The molecular basis for these differences is obscure. Spectrin is present throughout development, whereas band 3 is induced 2-3 days before enucleation. These developmental changes lead to the maturation of the cell membrane and cytoskeleton. The two- dimensional network of spectrin is reinforced by band 4.1 and short chains of actin. Spectrin is also linked to band 3 via ankyrin. This arrangement restricts the lateral mobility of the integral membrane protein. It is likely that the orderly assortment of these proteins noted with red cell development is influenced critically by their mutual interactions. This proposal will address the following: a) Does gelsolin, an actin-serving protein, regulate the length of F-actin in mammalian red cells, and does perturbation of actin polymerization interfere with protein sorting or cytoskeletal stability? b) Is the segregation of band 3 with the mature red cell a consequence of its binding to the cytoskeleton, and does it play a role in the long-term stability of the cytoskeleton? Specifically, a survey of the following proteins will be made to assess their assortment during development in MEL and mouse bone marrow cells: actin, spectrin, band 4.1, ankyrin, band 3, and gelsolin. Next, to modulate actin polymerization in vivo, MEL cells will be transfected with appropriate gelsolin construct to augment or suppress actin- severing activity at different stages of differentiation. Analogous studies with band 3 will also be performed to modulate its expression at various stages of erythropoiesis. The effects of these manipulations on cell morphology, osmotic fragility, enucleation, and protein assortment will be evaluated. In addition to probing basic mechanisms of protein sorting, these studies should improve our understanding of hemolytic anemias by providing cell culture models for these disorders.

蛋白质分类的机制在 分化和哺乳动物红细胞生成可提供有价值的 研究这种现象的模型。 红细胞蛋白质谱 例如，频段3保留在成熟的网状细胞中，而 大多数表面蛋白与挤出核分离。 这 这些差异的分子基础是晦涩的。 光谱是 在整个开发中存在，而频段3则诱导2-3天 摘除之前。 这些发展变化导致 细胞膜和细胞骨架的成熟。 两个 - 光谱的尺寸网络由频段4.1加强和短 肌动蛋白的链。 Spectrin还通过Ankyrin链接到Band 3。 这种布置限制了积分的横向迁移率 膜蛋白。 可能有序的 这些随红细胞发育所指出的蛋白质受到影响 批判性地通过他们的相互作用。 该提议将 地址以下： 调节哺乳动物红细胞中F-肌动蛋白的长度，并且DO 肌动蛋白聚合的扰动会干扰蛋白质排序 还是细胞骨架稳定性？ b）是频段3的隔离 成熟的红细胞与其与 细胞骨架，并且在长期稳定性中起作用 细胞骨架？ 具体而言，以下调查 将制定蛋白质来评估其开发过程中的分类 在MEL和小鼠骨髓细胞中：肌动蛋白，光谱，带4.1， Ankyrin，Band 3和Gelsolin。 接下来，调节肌动蛋白 在体内聚合，MEL细胞将被转染 适当的凝胶蛋白构造以增加或抑制肌动蛋白 - 分化的不同阶段的活动切断。 还将对3个带3进行类似研究以调节 它在红细胞生成的各个阶段的表达。 效果 这些关于细胞形态的操纵，渗透脆弱的， 将评估摘除剂和蛋白质分类。 除了探测蛋白质分类的基本机制外， 研究应通过 为这些疾病提供细胞培养模型。

项目成果

Structure and transcription of the mouse erythropoietin receptor gene.

小鼠促红细胞生成素受体基因的结构和转录。

• DOI: 10.1128/mcb.10.7.3675-3682.1990
• 发表时间: 1990
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Youssoufian,H;Zon,LI;Orkin,SH;D'Andrea,AD;Lodish,HF
• 通讯作者: Lodish,HF

Transcriptional inhibition of the murine erythropoietin receptor gene by an upstream repetitive element.

上游重复元件对小鼠促红细胞生成素受体基因的转录抑制。

• DOI: 10.1128/mcb.13.1.98-104.1993
• 发表时间: 1993
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Youssoufian,H;Lodish,HF
• 通讯作者: Lodish,HF

Further characterization of cis-acting regulatory sequences in the genomic locus of the murine erythropoietin receptor: evidence for stage-specific regulation.

鼠促红细胞生成素受体基因座中顺式作用调节序列的进一步表征：阶段特异性调节的证据。

• 发表时间: 1994
• 期刊: Blood
• 影响因子: 20.3
• 作者: Youssoufian,H

The cytoplasmic region of the erythropoietin receptor contains nonoverlapping positive and negative growth-regulatory domains.

促红细胞生成素受体的细胞质区域包含不重叠的正向和负向生长调节结构域。

• DOI: 10.1128/mcb.11.4.1980-1987.1991
• 发表时间: 1991
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: D'Andrea,AD;Yoshimura,A;Youssoufian,H;Zon,LI;Koo,JW;Lodish,HF
• 通讯作者: Lodish,HF

Activation of the erythropoietin receptor promoter by transcription factor GATA-1.

• DOI: 10.1073/pnas.88.23.10638
• 发表时间: 1991-12
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Leonard 1. Zon;Hagop YOUSSOUFIANtt;C. Mather;Harvey F. LODISHt;Stuart H. Orkin