The nuclear lamins A, B, and C are intermediate filament proteins that form a nuclear scaffold adjacent to the inner nuclear membrane in higher eukaryotes, providing structural support for the nucleus. In the past two decades it has become evident that the final step in the biogenesis of the mature lamin A from its precursor prelamin A by the zinc metalloprotease ZMPSTE24 plays a critical role in human health. Defects in prelamin A processing by ZMPSTE24 result in premature aging disorders including Hutchinson Gilford Progeria Syndrome (HGPS) and related progeroid diseases. Additional evidence suggests that defects in prelamin A processing, due to diminished ZMPSTE24 expression or activity, may also drive normal physiological aging. Because of the important connection between prelamin A processing and human aging, there is increasing interest in how ZMPSTE24 specifically recognizes and cleaves its substrate prelamin A, encoded by LMNA. Here, we describe two humanized yeast systems we have recently developed to examine ZMPSTE24 processing of prelamin A. These systems differ from one another slightly. Version 1.0 is optimized to analyze ZMPSTE24 mutations, including disease alleles that may affect the function or stability of the protease. Using this system, we previously showed that some ZMPSTE24 disease alleles that affect stability can be rescued by the proteasome inhibitor bortezomib, which may have therapeutic implications. Version 2.0 is designed to analyze LMNA mutations at or near the ZMPSTE24 processing site to assess whether they permit or impede prelamin A processing. Together these systems offer powerful methodology to study ZMPSTE24 disease alleles and to dissect the specific residues and features of the lamin A tail that are required for recognition and cleavage by the ZMPSTE24 protease.

中文翻译：

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一种人源化酵母系统，用于分析 ZMPSTE24 对 Prelamin A 的裂解

核纤层蛋白 A、B 和 C 是中间丝蛋白，它们在高等真核生物中形成与内核膜相邻的核支架，为细胞核提供结构支持。在过去的二十年中，很明显，锌金属蛋白酶 ZMPSTE24 从其前体 prelamin A 生成成熟 lamin A 的最后一步对人类健康起着至关重要的作用。ZMPSTE24 处理的 prelamin A 中的缺陷会导致过早衰老障碍，包括哈钦森吉尔福德早衰综合症 (HGPS) 和相关的早衰症。其他证据表明，由于 ZMPSTE24 表达或活性降低，prelamin A 加工中的缺陷也可能导致正常的生理老化。由于prelamin A加工与人类衰老之间的重要联系，人们越来越关注 ZMPSTE24 如何特异性识别和切割其由 LMNA 编码的底物前核蛋白 A。在这里，我们描述了我们最近开发的两个人源化酵母系统，用于检查预蛋白 A 的 ZMPSTE24 加工。这些系统彼此略有不同。1.0 版经过优化，可分析 ZMPSTE24 突变，包括可能影响蛋白酶功能或稳定性的疾病等位基因。使用该系统，我们之前表明，蛋白酶体抑制剂硼替佐米可以挽救一些影响稳定性的 ZMPSTE24 疾病等位基因，这可能具有治疗意义。2.0 版旨在分析 ZMPSTE24 加工位点处或附近的 LMNA 突变，以评估它们是否允许或阻碍 prelamin A 加工。