文献类型：期刊文献

英文题名：Nucleotide distribution analysis of 5′UTRs in genome-scale directs their redesign and expression regulation in yeast

作者：Yao, Chaoying[1];Yin, Yu[2];Li, Qingyang[1];Zhang, Hanqi[1];Zhang, Yilun[1];Shao, Qianqian[1];Liu, Qi[1];Ren, Yanna[1];Cai, Menghao[1,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Sch Pharm, Lab Pharmaceut Crystal Engn & Technol, 130 Meilong Rd, Shanghai 200237, Peoples R China;[3]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2025

卷号：88

起止页码：113

外文期刊名：METABOLIC ENGINEERING

收录：;EI(收录号：20250117622134);WOS:【SCI-EXPANDED(收录号:WOS:001421091200001)】；

基金：Funding This study was supported by the National Key R & D Program of China (2022YFC2805102) , National Natural Science Foundation of China (32201206) , Shanghai Explorer Program (24TS1411600) and China Postdoctoral Science Foundation (2022M711146) .

语种：英文

外文关键词：Gene expression regulation - Nucleotides - Transcription

摘要：Non-conventional yeasts have emerged as important sources of valuable products in bioindustries. However, tools for the control of expression are limited in these hosts. In this study, we aimed to excavate the tools for the regulation of translation that are often overlooked. 5 ' UTR analysis of genome-scale annotated genes of four yeast species revealed a distinct decreasing 'G' frequency in -100 similar to -1 region from 5040 5 ' UTRs in Komagataella phaffii. New 5 ' UTRs were regenerated by base substitutions in defined regions, and replacement of 'G' by 'A' or 'T' in the -50 similar to -1 region highly facilitated gene expression. Preference analysis of all nucleotide triplets in 5 ' UTRs revealed a KZ(3) (-3 similar to -1) that dominantly affected gene expression. A total of 128 KZ(3) variants were constructed to work with promoters of methanol-inducible P-AOX1 and constitutive P-GAP, of which 58 KZ(3) variants increased gene expression and maximum difference in strength was 15-fold among all variants. Polysome profiling analysis clarified that 5 ' UTR-KZ(3) enhanced gene expression at translational but not transcriptional levels. Finally, improved production of three industrial proteins and one platform compound were achieved by ready-made 5 ' UTR-KZ(3) or in situ modification of the 5 ' UTR. This study provides new references and tools for the fine-tuning of translational regulation in yeast and other fungi.