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Due-B对于脊椎动物的早期发育和基因组复制是可有可无的。

Due-B Is dispensable for early development and genome duplication in vertebrates.

作者信息

Sansam Courtney G, Masser Emily A, Goins Duane, Sansam Christopher L

机构信息

Oklahoma Medical Research Foundation, Cell Cycle and Cancer Biology Research Program, Oklahoma City, OK USA.

University of Oklahoma Health Sciences Center, Department of Cell Biology, Oklahoma City, OK USA.

出版信息

bioRxiv. 2025 Jun 17:2025.06.17.658125. doi: 10.1101/2025.06.17.658125.

Abstract

The DNA Unwinding Element-Binding protein (DUE-B) is a Cyclin Dependent Kinase (CDK) and Dbf4-Dependent Kinase (DDK) substrate that has been implicated in the control of DNA replication initiation. Knockdown of DUE-B in HeLa cells perturbs the G1-to-S phase transition and immunodepletion of in egg extracts blocks replication initiation. Combined prior evidence has suggested that may be a vertebrate-specific DNA replication initiation factor. Here, we asked whether was an essential vertebrate gene , and whether it was critical for proper embryonic development in the zebrafish . We have generated mutant zebrafish through genome-editing TALENs that fail to express mRNA or protein. Our mutant zebrafish are viable and survive to adulthood. They do not display outward developmental phenotypes, and when stressed with replication inhibitors, do not differ from their wild-type counterparts. Cell cycle analysis demonstrates that DNA replication occurs normally. Our data indicate that Due-b is not a core essential component of the DNA replication initiation machinery required for vertebrate development. Instead, it may be a mechanistically redundant protein or play a specialized role in DNA replication control.

摘要

DNA解旋元件结合蛋白(DUE-B)是一种细胞周期蛋白依赖性激酶(CDK)和Dbf4依赖性激酶(DDK)底物,与DNA复制起始的控制有关。在HeLa细胞中敲低DUE-B会扰乱G1期到S期的转换,在卵提取物中对其进行免疫去除会阻断复制起始。先前的综合证据表明,DUE-B可能是脊椎动物特有的DNA复制起始因子。在这里,我们研究了DUE-B是否是脊椎动物必需基因,以及它对斑马鱼正常胚胎发育是否至关重要。我们通过基因组编辑TALENs产生了不表达DUE-B mRNA或蛋白质的突变斑马鱼。我们的突变斑马鱼是有活力的,能存活到成年。它们没有表现出外在的发育表型,在用复制抑制剂处理时,与野生型斑马鱼没有差异。细胞周期分析表明DNA复制正常发生。我们的数据表明,DUE-B不是脊椎动物发育所需的DNA复制起始机制的核心必需成分。相反,它可能是一种机制上冗余的蛋白质,或在DNA复制控制中发挥特殊作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5265/12262730/29ffa5e5cfc3/nihpp-2025.06.17.658125v1-f0001.jpg

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