The role of the human transcription factor ZFP64 in centromeric transcription and maintenance of genomic stability

人类转录因子ZFP64在着丝粒转录和维持基因组稳定性中的作用

• 批准号: BB/V009605/1
• 负责人: Sarah McClelland
• 金额: $ 55.22万
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV009605%2F1
• 关键词: role; human; transcription; factor; ZFP64

Our bodies are comprised of 30 billion cells. As our body grows, and regenerates tissues due to injury, normal wear-and-tear and aging, these cells must multiply at rapid rates - with millions of new cells born every minute in our bodies! Each time a cell duplicates its content, including its copy of the genome, it undergoes a process termed cell division that physically separates the cell contents into two halves that become the future daughter cells. Any mistakes that occur in the precise separation of the two new genomes can lead to dramatic and potentially cancer-causing genetic mutations, or cell death. As a result, cell division is carefully regulated by many control mechanisms. The genome is arranged into 22 pairs of chromosomes that, once copied, are plucked apart by internal cell structures to form the genome of the new cells. During this process (mitosis), chromosomes are captured by the cell via specialised structures called centromeres. Most of the genome is relentlessly read and 'transcribed' by proteins that create an RNA copy of short sections - usually as a precursor to producing proteins from the genes encoded in our DNA. Centromeres were thought to be excluded from this process but over the last several years it was surprisingly discovered that transcription of centromeres is essential to accurate cell division.This crucially important and evolutionarily-conserved process has been difficult to study because of two major hurdles; First, it is not known which exact DNA sequences undergo transcription. Second, the specific proteins (transcription factors) that perform this role at the centromere are not known. Recently, our laboratory has identified a transcription factor named ZFP64 as a potential centromeric transcription factor. We used imaging to track the position of this factor within human cells, and excitingly see it at centromeres. Importantly, when we specifically remove this protein from cells we see a decrease in the amount of centromeric transcription. We have also discovered the possible mechanism by which centromeric transcription is controlled throughout the lifetime of each cell - another elusive phenomenon in humans. Therefore, we are now poised to take advantage of recent technological advances that will improve our ability to discover which DNA sequences at the centromere are transcribed, and to ask which of those are controlled by ZFP64. Our laboratory is expert in imaging and manipulating human cells and we have assembled a focussed and feasible set of experiments that will allow us to definitively test whether ZFP64 indeed represents the first human centromeric transcription factor. Together with expert collaborators within our research institute and worldwide, we will perform both standard experiments and also newly emerging strategies for discovering which DNA sequences are transcribed. By investigating the detailed mechanisms of this new and important potential centromeric transcription factor, we will make a large step towards full understanding of this phenomenon in humans, and exactly why it is so crucial for accurate cell division.

我们的身体由300亿个细胞组成。随着我们身体的生长，由于受伤，正常的磨损和衰老而再生组织，这些细胞必须以快速的速度繁殖-每分钟都有数百万个新细胞在我们体内诞生！每次细胞复制其内容物，包括其基因组拷贝时，它都会经历一个称为细胞分裂的过程，该过程将细胞内容物物理分离成两半，成为未来的子细胞。在两个新基因组的精确分离中发生的任何错误都可能导致戏剧性的和潜在的致癌基因突变或细胞死亡。因此，细胞分裂受到许多控制机制的精心调控。基因组被排列成22对染色体，一旦被复制，就会被内部细胞结构分开，形成新细胞的基因组。在这个过程（有丝分裂）中，染色体通过称为着丝粒的特殊结构被细胞捕获。基因组的大部分都被蛋白质无情地读取和“转录”，这些蛋白质产生了一个短片段的RNA拷贝--通常是从我们DNA中编码的基因中产生蛋白质的前体。着丝粒被认为是排除在这一过程之外的，但在过去的几年里，人们惊讶地发现，着丝粒的转录对精确的细胞分裂是必不可少的。这一至关重要的和进化上保守的过程一直很难研究，因为有两个主要的障碍：首先，不知道确切的DNA序列进行转录。第二，在着丝粒处起这种作用的特定蛋白质（转录因子）尚不清楚。最近，我们实验室鉴定了一个名为ZFP 64的转录因子作为潜在的着丝粒转录因子。我们使用成像技术来追踪这种因子在人类细胞中的位置，并令人兴奋地看到它在着丝粒上。重要的是，当我们从细胞中特异性地去除这种蛋白质时，我们看到着丝粒转录的量减少。我们还发现了在每个细胞的整个生命周期中控制着丝粒转录的可能机制-这是人类中另一种难以捉摸的现象。因此，我们现在准备利用最新的技术进步，这将提高我们发现着丝粒上哪些DNA序列被转录的能力，并询问哪些DNA序列由ZFP 64控制。我们的实验室是成像和操纵人类细胞的专家，我们已经组装了一组集中和可行的实验，这将使我们能够明确测试ZFP 64是否确实代表第一个人类着丝粒转录因子。与我们研究所和世界各地的专家合作者一起，我们将进行标准实验和新出现的策略，以发现哪些DNA序列被转录。通过研究这种新的和重要的潜在的着丝粒转录因子的详细机制，我们将朝着充分理解人类的这种现象迈出一大步，以及为什么它对准确的细胞分裂如此重要。

项目成果

Replication stress generates distinctive landscapes of DNA copy number alterations and chromosome scale losses.

• DOI: 10.1186/s13059-022-02781-0
• 发表时间: 2022-10-20
• 期刊: Genome biology
• 影响因子: 12.3

Additional file 5 of Replication stress generates distinctive landscapes of DNA copy number alterations and chromosome scale losses

复制压力的附加文件 5 会产生 DNA 拷贝数改变和染色体规模损失的独特景观

• DOI: 10.6084/m9.figshare.21376248
• 发表时间: 2022
• 作者: Shaikh N

Targeted assembly of ectopic kinetochores to induce chromosome-specific segmental aneuploidies.

• DOI: 10.15252/embj.2022111587
• 发表时间: 2023-05-15
• 期刊: The EMBO journal

Inducing Specific Chromosome Mis-Segregation in Human Cells

诱导人体细胞中的特定染色体错误分离

• DOI: 10.1101/2022.04.19.486691
• 发表时间: 2022
• 作者: Tovini L

Additional file 1 of Replication stress generates distinctive landscapes of DNA copy number alterations and chromosome scale losses

复制压力的附加文件 1 会产生 DNA 拷贝数改变和染色体规模损失的独特景观

• DOI: 10.6084/m9.figshare.21376236
• 发表时间: 2022
• 作者: Shaikh N