Regulation of proliferation and differentiation in the male germ line adult stem cell lineage

雄性生殖系成体干细胞谱系增殖和分化的调节

基本信息

批准号：
10675340
负责人：
MARGARET T FULLER
金额：
$ 1.41万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2025-05-31
项目状态：
未结题

项目摘要

PROJECT SUMMARY (of Parent Project): Regulation of proliferation and differentiation in the male germ line adult stem cell lineage The switch from proliferation to differentiation is a key regulatory point in both embryonic development and the adult stem cell lineages that underlie tissue maintenance and repair. Failure of this switch may contribute to genesis of cancer. My laboratory has long used the Drosophila male germ line as a model to investigate how self-renewal, proliferation and differentiation are regulated in adult stem cell lineages. Several lines of our inquiry have recently begun to converge on the molecular mechanisms underlying the developmentally programmed transition from mitotic proliferation to onset of meiosis and differentiation, implicating a number of molecular and cellular mechanisms in regulating this critical switch. We find that RNA binding proteins involved in translational control and alternative splicing act cell autonomously to regulate the cessation of proliferation and that progression of differentiation requires communication from associated somatic support cells. We discovered that a developmentally regulated switch in the site at which specific nascent transcripts are cut to form 3’ ends, leading to production of novel mRNA isoforms with shortened 3’UTRs, controls dramatic changes in the suite of proteins expressed in differentiating spermatocytes compared to proliferating spermatogonia. We found that dramatic changes in chromatin open over 2000 new promoters with novel core sequence structure to turn on the new cell type specific transcription program when cells initiate spermatocyte differentiation. Some of the earliest genes turned on in this differentiation program encode chromatin associated proteins that prevent spurious opening of normally cryptic promoters, thus preventing massive misexpression of genes associated with the wrong cell type. Other transcripts upregulated with differentiation onset encode cell type-specific translational regulators that delay production of core G2/M cell cycle machinery to program the extended G2 phase of meiotic prophase. Over the next 5 years, we propose to map how these processes collaborate to form the regulatory circuitry that initiates then executes the switch from mitosis to meiosis. We will investigate how the RNA binding proteins Bam and Bgcn trigger the switch from mitosis to differentiation by repressing expression of the alternative splice factor HOW, identify candidate substrates of HOW by immunoprecipitation followed by RNA- Seq, and assess their function in vivo, including whether they communicate with adjacent somatic support cells. We will investigate how the switch in proteins expressed due to alternative 3’ end cut site selection on nascent transcripts is regulated and influences differentiation. We will investigate how cell-type specific chromatin regulators and proteins that recruit them to specific loci set up the new transcription program for differentiation. To elucidate how the developmental program remodels fundamental cellular processes like the cell cycle to set up differentiation of specialized cell types, we will investigate how cell-type specific RNA binding proteins first repress, then activate translation of cyclin B during meiotic prophase and how the DAZ homolog Boule regulates progression into the meiotic divisions.

项目摘要（父项目）：雄性生殖系成体干细胞谱系增殖和分化的调节从增殖到分化的转变是胚胎发育和胚胎发育的关键调节点。作为组织维持和修复基础的成体干细胞谱系的失败可能会导致这种转变。我的实验室长期以来一直使用果蝇雄性生殖系作为模型来研究癌症的起源。自我更新、增殖和分化在成体干细胞谱系中受到调节。最近开始集中于发育程序背后的分子机制从有丝分裂增殖到减数分裂和分化开始的转变，涉及许多分子和我们发现 RNA 结合蛋白参与翻译。控制和选择性剪接作用细胞自主地调节增殖的停止，并且我们发现分化的进展需要相关体细胞支持细胞的通信。特定新生转录物被切割形成 3' 末端的位点中的发育调节开关，导致产生具有缩短的 3'UTR 的新型 mRNA 亚型，控制一系列的显着变化与增殖的精原细胞相比，分化的精母细胞中表达的蛋白质。染色质的巨大变化打开了 2000 多个具有新颖核心序列结构的新启动子，以开启当细胞启动精母细胞分化时，新的细胞类型特异性转录程序。在这个分化程序中打开的基因编码染色质相关蛋白，以防止虚假打开通常隐藏的启动子，从而防止与错误的细胞类型。分化开始时上调的其他转录物编码细胞类型特异性翻译。延迟核心 G2/M 细胞周期机制产生的调节因子，以编程延长减数分裂的 G2 阶段在接下来的 5 年里，我们建议绘制这些流程如何协作形成监管的图景。启动然后执行从有丝分裂到减数分裂的转换的电路我们将研究 RNA 是如何结合的。 Bam 和 Bgcn 蛋白通过抑制替代剪接因子 HOW，通过免疫沉淀和 RNA- 鉴定 HOW 的候选底物测序，并评估它们的体内功能，包括它们是否与相邻的体细胞支持细胞通讯。我们将研究新生蛋白中由于替代 3' 末端切割位点选择而导致的蛋白质转换如何表达。我们将研究细胞类型特异性染色质如何受到调节并影响分化。将它们招募到特定位点的调节因子和蛋白质建立了新的分化转录程序。阐明发育程序如何重塑细胞周期等基本细胞过程以设定为了提高特殊细胞类型的分化，我们将首先研究细胞类型特异性 RNA 结合蛋白如何在减数分裂前期抑制并激活细胞周期蛋白 B 的翻译以及 DAZ 同源物 Boule 如何调节进展到减数分裂。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Developmental Regulation of Alternative Polyadenylation in an Adult Stem Cell Lineage.

成体干细胞谱系中替代多腺苷酸化的发育调节。

DOI：
10.1101/2024.03.18.585561
发表时间：
2024
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Gallicchio,Lorenzo;Matias,NeuzaR;Morales-Polanco,Fabian;Nava,Iliana;Stern,Sarah;Zeng,Yi;Fuller,MargaretT
通讯作者：
Fuller,MargaretT

Regulation and function of alternative polyadenylation in development and differentiation.

DOI：
10.1080/15476286.2023.2275109
发表时间：
2023-01
期刊：
RNA BIOLOGY
影响因子：
4.1
作者：
Gallicchio, Lorenzo;Olivares, Gonzalo H.;Berry, Cameron W.;Fuller, Margaret T.
通讯作者：
Fuller, Margaret T.

Identification of Protein-RNA Interactions in Mouse Testis Tissue Using fRIP.

使用 fRIP 鉴定小鼠睾丸组织中的蛋白质-RNA 相互作用。

DOI：
10.21769/bioprotoc.4286
发表时间：
2022
期刊：
Bio-protocol
影响因子：
0.8
作者：
Bailey,AlexisS;Batista,PedroJ;Chang,HowardY;Fuller,MargaretT
通讯作者：
Fuller,MargaretT

Developmentally regulated alternate 3' end cleavage of nascent transcripts controls dynamic changes in protein expression in an adult stem cell lineage.

DOI：
10.1101/gad.349689.122
发表时间：
2022-08-01
期刊：
GENES & DEVELOPMENT
影响因子：
10.5
作者：
Berry, Cameron W;Olivares, Gonzalo H;Gallicchio, Lorenzo;Ramaswami, Gokul;Glavic, Alvaro;Olguin, Patricio;Li, Jin Billy;Fuller, Margaret T
通讯作者：
Fuller, Margaret T