Species-specific protein-protein interactions govern the humanization of the 20S proteasome in yeast.
Species-specific protein-protein interactions govern the humanization of the 20S proteasome in yeast.
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DOI：
10.1093/genetics/iyad117
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发表时间：
2023-08-31
期刊：
影响因子：
3.3
通讯作者：
Kachroo, Aashiq H.
中科院分区：
文献类型：
--
作者：
Sultana, Sarmin;Abdullah, Mudabir;Li, Jianhui;Hochstrasser, Mark;Kachroo, Aashiq H.
关键词：
酵母和人类共享数千个基因，尽管有十亿年的进化分歧。虽然许多人类基因可以在功能上取代它们的酵母对应物，但近一半的测试共享基因不能。例如，大多数酵母蛋白酶体亚基是“可人源化的”，除了包含β-环核心的亚基，包括β2c（HsPSMB 7，组成型蛋白酶体亚基）。我们开发了一个高通量的管道，通过产生一个大型的Hsβ2c突变体库，并筛选它们的酵母β2（ScPup 1）敲除的互补来人源化酵母蛋白酶体。能够取代ScPup 1的变体包括（1）影响局部蛋白质-蛋白质相互作用（PPI）的变体，其中最影响β2c C-末端尾和相邻β3亚基之间的相互作用，以及（2）影响β2c蛋白水解活性的变体。将人β2c的全长尾与ScPup 1的全长尾交换使互补成为可能。此外，如果还提供人β3，则野生型人β2c可以替代酵母β2。出乎意料的是，酵母蛋白酶体携带催化失活的HsPSMB 7-T44 A变体，阻断前体自动加工是可行的，这表明一个完整的前肽稳定后期组装中间体。相反，人β 2 i（HsPSMB 10）（一种免疫蛋白酶体亚基和酵母β2的共直系同源物）中的类似修饰不能在酵母中实现互补，表明人免疫蛋白酶体核心组装中涉及不同的相互作用。从广义上讲，我们的数据揭示了特定PPI在巨大的进化距离上管理功能可替换性的作用。
Yeast and humans share thousands of genes despite a billion years of evolutionary divergence. While many human genes can functionally replace their yeast counterparts, nearly half of the tested shared genes cannot. For example, most yeast proteasome subunits are “humanizable,” except subunits comprising the β-ring core, including β2c (HsPSMB7, a constitutive proteasome subunit). We developed a high-throughput pipeline to humanize yeast proteasomes by generating a large library of Hsβ2c mutants and screening them for complementation of a yeast β2 (ScPup1) knockout. Variants capable of replacing ScPup1 included (1) those impacting local protein–protein interactions (PPIs), with most affecting interactions between the β2c C-terminal tail and the adjacent β3 subunit, and (2) those affecting β2c proteolytic activity. Exchanging the full-length tail of human β2c with that of ScPup1 enabled complementation. Moreover, wild-type human β2c could replace yeast β2 if human β3 was also provided. Unexpectedly, yeast proteasomes bearing a catalytically inactive HsPSMB7-T44A variant that blocked precursor autoprocessing were viable, suggesting an intact propeptide stabilizes late assembly intermediates. In contrast, similar modifications in human β2i (HsPSMB10), an immunoproteasome subunit and the co-ortholog of yeast β2, do not enable complementation in yeast, suggesting distinct interactions are involved in human immunoproteasome core assembly. Broadly, our data reveal roles for specific PPIs governing functional replaceability across vast evolutionary distances.
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影响因子：
--
作者：
Kuzmin, Elena;Costanzo, Michael;Boone, Charles
通讯作者：
Boone, Charles
影响因子：
5.7
作者：
Harshbarger, Wayne;Miller, Chase;Sacchettini, James
通讯作者：
Sacchettini, James
影响因子：
0.8
作者：
Akhmetov, Azat;Laurent, Jon M.;Marcotte, Edward M.
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Marcotte, Edward M.
DOI：
10.1016/j.crmeth.2023.100464
发表时间：
2023-05-22
期刊：
Cell reports methods
影响因子：
--
作者：
通讯作者：
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影响因子：
16.8
作者：
Kusmierczyk, Andrew R.;Kunjappu, Mary J.;Hochstrasser, Mark
通讯作者：
Hochstrasser, Mark