Chromatin Expulsion by the DNA Replication Licensing Factor ORC4 during Asymmetric Cell Division in Meiosis and Differentiation

减数分裂和分化过程中不对称细胞分裂过程中 DNA 复制许可因子 ORC4 的染色质排出

基本信息

批准号：
10059254
负责人：
WILLIAM S WARD
金额：
$ 31.84万
依托单位：
UNIVERSITY OF HAWAII AT MANOA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2022-11-30
项目状态：
已结题

项目摘要

The goal of studies described in this amended R01 application is to explore the role of the DNA licensing factor, ORC4, in sequestering chromosomes into polar bodies via cage formation during female meiosis. These studies were considered highly innovative and with fundamental significance for understanding the mechanism of female meiotic division and oocyte maturation, as well as other possible forms of asymmetric cell division. Other strengths of the application include the outstanding qualifications and record of productivity of the Principal Investigator, the recruitment of a strong collaborative team with complementary expertise, the highly conducive research environment at the University of Hawaii, the convincing premise in the form of previous publications and preliminary data confirming the underlying functionalities of ORC4, the availability of many key reagents and resources required to pursue the project goals, adequate attention to elements of scientific rigor and a strong response to the previous critiques. In addition to these abundant strengths, some expressed concerns with an underdeveloped approach, including lack of guidance as to the mechanistic implications for experiments involving ORC4 depletion, beyond previous observations, concerns that overexpression of the EGFP-ORC-4 fusion protein may induce cell stress or lethality, lack of feasibility of purification of ORC4 interacting partners and the absence of sufficient insight on the role of ORC4 in other forms of asymmetric cell division. While these and other minor weaknesses slightly reduced the perceived merit of the application, the panel ultimately concluded that the very high novelty and fundamental mechanistic insights to be gained will exert a major impact on the fields of oocyte development and developmental biology. rtility.

修订后的 R01 申请中描述的研究目标是探索 DNA 许可因子 ORC4 在雌性减数分裂期间通过笼形成将染色体隔离到极体中的作用。这些研究被认为具有高度创新性，对于理解雌性减数分裂和卵母细胞成熟的机制以及其他可能形式的不对称细胞分裂具有根本意义。申请的其他优势包括首席研究员的杰出资质和生产力记录、招募具有互补专业知识的强大协作团队、夏威夷大学高度有利的研究环境、以前出版物形式的令人信服的前提和初步数据确认了 ORC4 的基本功能、实现项目目标所需的许多关键试剂和资源的可用性、对科学严谨性要素的充分关注以及对先前批评的强烈回应。除了这些丰富的优势之外，一些人还对不发达的方法表示担忧，包括缺乏对涉及 ORC4 耗竭的实验的机制影响的指导，超出了之前的观察结果，担心 EGFP-ORC-4 融合蛋白的过度表达可能会诱发细胞应激或致死率、缺乏纯化 ORC4 相互作用伴侣的可行性以及对 ORC4 在其他形式的不对称细胞分裂中的作用缺乏足够的了解。虽然这些和其他小缺陷稍微降低了该应用的感知优点，但专家组最终得出的结论是，获得的非常高的新颖性和基本机制见解将对卵母细胞发育和发育生物学领域产生重大影响。实用性。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

AKAP13 couples GPCR signaling to mTORC1 inhibition.

AKAP13 将 GPCR 信号传导与 mTORC1 抑制结合起来。

DOI：
发表时间：
2021
期刊：
影响因子：
4.5
作者：
Zhang, Shihai;Wang, Huanyu;Melick, Chase H;Jeong, Mi;Curukovic, Adna;Tiwary, Shweta;Lama;Meng, Delong;Servage, Kelly A;James, Nicholas G;Jewell, Jenna L
通讯作者：
Jewell, Jenna L

Sperm degradation after vasectomy follows a sperm chromatin fragmentation-dependent mechanism causing DNA breaks in the toroid linker regions.

输精管切除术后的精子降解遵循精子染色质碎片依赖性机制，导致环形接头区域 DNA 断裂。

DOI：
发表时间：
2022-08-29
期刊：
影响因子：
4
作者：
Ribas;Nguyen, Hieu;Valle, Raquel;Wu, Hongwen;Yeste, Marc;Ward, W Steven
通讯作者：
Ward, W Steven

Spatial and temporal resolution of mORC4 fluorescent variants reveals structural requirements for achieving higher order self-association and pronuclei entry.

mORC4 荧光变体的空间和时间分辨率揭示了实现更高阶自缔合和原核进入的结构要求。

DOI：
发表时间：
2019-05-16
期刊：
影响因子：
0
作者：
Nguyen, Hieu;Ward, W Steven;James, Nicholas G
通讯作者：
James, Nicholas G

ArfGAP1 inhibits mTORC1 lysosomal localization and activation.

ArfGAP1 抑制 mTORC1 溶酶体定位和激活。

DOI：
发表时间：
2021
期刊：
影响因子：
0
作者：
Meng, Delong;Yang, Qianmei;Melick, Chase H;Park, Brenden C;Hsieh, Ting;Curukovic, Adna;Jeong, Mi;Zhang, Junmei;James, Nicholas G;Jewell, Jenna L
通讯作者：
Jewell, Jenna L

The role of ORC4 in enucleation of Murine Erythroleukemia (MEL) cells is similar to that in oocyte polar body extrusion.

ORC4 在小鼠红白血病 (MEL) 细胞去核中的作用与在卵母细胞极体排出中的作用相似。