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酵母染色体组合因子1与增殖细胞核抗原之间的相互作用的结构基础

Keely S Orndorff¹, Evan J Veltri¹, Nicole M Hoitsma²

¹Department of Chemistry and Biochemistry, Creighton University, Omaha, NE, USA.

Journal of molecular biology

|July 5, 2024

概括

增殖细胞核抗原 (PCNA) 通过特定的动机与染色体组合因子1 (CAF-1) 相互作用. 带正电荷的侧面氨基酸增强了这一关键的DNA复制相互作用.

科学领域:

分子生物学分子生物学
结构生物学结构生物学
生物化学生物化学

背景情况:

增殖细胞核抗原 (PCNA) 是DNA复制必不可少的同位三元蛋白,在复制分叉上协调蛋白质活动.
染色体组装因子1 (CAF-1) 是一个基因组伴侣,它在DNA复制后沉积基因组,并与PCNA相互作用,在沉默区域进行核细胞组装.
PCNA通过短PCNA相互作用 (PIP) 动机歧视结合伙伴的确切机制尚不清楚.

研究的目的:

阐明酵母CAF-1 PIP动机与PCNA之间的相互作用的结构基础.
调查侧面氨基酸和电荷分布在调节CAF-1与PCNA的结合亲和力中的作用.

主要方法:

酵母CAF-1 PIP动机结合PCNA的晶体结构的确定.
采用了一种新的策略,用于PCNA-PIP复合体的静止学生产.
进行突变分析以评估侧面氨基酸电荷对结合亲和力的影响.

主要成果:

晶体结构显示,CAF-1 PIP动机与PCNA上的疏水口袋结合,与已知的相互作用一致.
围绕PIP动机的氨基酸与PCNA IDCL或C端相互作用,这种特征在其他有限的相互作用中观察到.
突变分析表明,侧边区域的正电荷对于低微分子亲和力至关重要,而上游的负电荷则降低了结合.

结论:

CAF-1和PCNA之间的相互作用不仅涉及正规的PIP动机,还涉及相邻的区域.
侧面区域的正电荷对于PCNA相互作用蛋白的特异性和亲和性至关重要.
这些发现提供了关于PCNA如何在复制分叉上招募蛋白质的特异性的见解.

关键词:

染色体组合因子1 染色体组合因子1 基因沉默是一种基因沉默.核组合组合的核组合.与PCNA相互作用的 (PIP)增殖细胞核抗原的核抗原.

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