TY - JOUR

T1 - A Kinase and a Glycosylase Catabolize Pseudouridine in the Peroxisome to Prevent Toxic Pseudouridine Monophosphate Accumulation

AU - Chen, Mingjia

AU - Witte, Claus-Peter

N1 - Funding Information: We thank Anting Zhu for preparing vector V89, André Specht, Hildegard Thölke for technical support, Katharina J. Heinemann and Xiaoguang Chen for help with plant cultivation and harvesting, Xiaoye Liu for assistance with the statistical analyses, Wenlei Wang for help with salt stress analyses, and Nieves Medina Escobar for revising the article. This work was financially supported by the National Natural Science Foundation of China (grant 31900907), the Natural Science Foundation of Jiangsu Province (grant BK20190528), the Leibniz University Hannover (Wege in die Forschung II to M.C.), and Deutsche Forschungsgemeinschaft (grant CH2292/1-1 to M.C. and grant WI3411/4-1 to C.-P.W.)

PY - 2020/3

Y1 - 2020/3

N2 - Pseudouridine (C) is a frequent nucleoside modification that occurs in both noncoding RNAs and mRNAs. In pseudouridine, C5 of uracil is attached to the Rib via an unusual C-glycosidic bond. This RNA modification is introduced on the RNA by site-specific transglycosylation of uridine (U), a process mediated by pseudouridine synthases. RNA is subject to constant turnover, releasing free pseudouridine, but the metabolic fate of pseudouridine in eukaryotes is unclear. Here, we show that in Arabidopsis (Arabidopsis thaliana), pseudouridine is catabolized in the peroxisome by (1) a pseudouridine kinase (PUKI) from the PfkB family that generates 59-pseudouridine monophosphate (59-CMP) and (2) a CMP glycosylase (PUMY) that hydrolyzes CMP to uracil and ribose-5-phosphate. Compromising pseudouridine catabolism leads to strong pseudouridine accumulation and increased CMP content. CMP is toxic, causing delayed germination and growth inhibition, but compromising pseudouridine catabolism does not affect the C/U ratios in RNA. The bipartite peroxisomal PUKI and PUMY are conserved in plants and algae, whereas some fungi and most animals (except mammals) possess a PUMY-PUKI fusion protein, likely in mitochondria. We propose that vacuolar turnover of ribosomal RNA produces most of the pseudouridine pool via 39-CMP, which is imported through the cytosol into the peroxisomes for degradation by PUKI and PUMY, a process involving a toxic 59-CMP intermediate.

AB - Pseudouridine (C) is a frequent nucleoside modification that occurs in both noncoding RNAs and mRNAs. In pseudouridine, C5 of uracil is attached to the Rib via an unusual C-glycosidic bond. This RNA modification is introduced on the RNA by site-specific transglycosylation of uridine (U), a process mediated by pseudouridine synthases. RNA is subject to constant turnover, releasing free pseudouridine, but the metabolic fate of pseudouridine in eukaryotes is unclear. Here, we show that in Arabidopsis (Arabidopsis thaliana), pseudouridine is catabolized in the peroxisome by (1) a pseudouridine kinase (PUKI) from the PfkB family that generates 59-pseudouridine monophosphate (59-CMP) and (2) a CMP glycosylase (PUMY) that hydrolyzes CMP to uracil and ribose-5-phosphate. Compromising pseudouridine catabolism leads to strong pseudouridine accumulation and increased CMP content. CMP is toxic, causing delayed germination and growth inhibition, but compromising pseudouridine catabolism does not affect the C/U ratios in RNA. The bipartite peroxisomal PUKI and PUMY are conserved in plants and algae, whereas some fungi and most animals (except mammals) possess a PUMY-PUKI fusion protein, likely in mitochondria. We propose that vacuolar turnover of ribosomal RNA produces most of the pseudouridine pool via 39-CMP, which is imported through the cytosol into the peroxisomes for degradation by PUKI and PUMY, a process involving a toxic 59-CMP intermediate.

UR - http://www.scopus.com/inward/record.url?scp=85081139259&partnerID=8YFLogxK

U2 - 10.1105/tpc.19.00639

DO - 10.1105/tpc.19.00639

M3 - Article

VL - 32

SP - 722

EP - 739

JO - The plant cell

JF - The plant cell

SN - 1040-4651

IS - 3

ER -