Loss of the RNA trimethylguanosine cap is compatible with nuclear accumulation of spliceosomal snRNAs but not pre-mRNA splicing or snRNA processing during animal development

• Published in: PLoS genetics Vol. 16; no. 10; p. e1009098
• Main Authors: Cheng, Lin, Zhang, Yu, Zhang, Yi, Chen, Tao, Xu, Yong-Zhen, Rong, Yikang S.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 21.10.2020; Public Library of Science (PLoS)
• Subjects: Animal development; Animals; Biology and life sciences; Chemical properties; Chromosomes; Comparative analysis; Drosophila; Drosophila melanogaster - genetics; Enzymes; Genetic aspects; Guanosine - analogs & derivatives; Guanosine - genetics; Guanosine - metabolism; Identification and classification; Insects; Introns; Introns - genetics; Larva - genetics; Larva - growth & development; Lethality; Life sciences; Localization; Mammals; Methyltransferases - genetics; mRNA; Mutagenesis; Mutation; Non-coding RNA; Nuclear transport; Nucleoli; Organisms; Proteins; Research and Analysis Methods; RNA; RNA Caps - genetics; RNA splicing; RNA Splicing - genetics; RNA, Messenger - genetics; RNA, Small Nuclear - genetics; Saccharomyces cerevisiae - genetics; Sequence Analysis, RNA - methods; snRNA; Software; Spliceosomes; Spliceosomes - genetics; Splicing; Trimethylguanosine; China; United States > US
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1009098

The 2,2,7-trimethylguanosine (TMG) cap is one of the first identified modifications on eukaryotic RNAs. TMG, synthesized by the conserved Tgs1 enzyme, is abundantly present on snRNAs essential for pre-mRNA splicing. Results from ex vivo experiments in vertebrate cells suggested that TMG ensures nuclear localization of snRNAs. Functional studies of TMG using tgs1 mutations in unicellular organisms yield results inconsistent with TMG being indispensable for either nuclear import or splicing. Utilizing a hypomorphic tgs1 mutation in Drosophila, we show that TMG reduction impairs germline development by disrupting the processing, particularly of introns with smaller sizes and weaker splice sites. Unexpectedly, loss of TMG does not disrupt snRNAs localization to the nucleus, disputing an essential role of TMG in snRNA transport. Tgs1 loss also leads to defective 3' processing of snRNAs. Remarkably, stronger tgs1 mutations cause lethality without severely disrupting splicing, likely due to the preponderance of TMG-capped snRNPs. Tgs1, a predominantly nucleolar protein in Drosophila, likely carries out splicing-independent functions indispensable for animal development. Taken together, our results suggest that nuclear import is not a conserved function of TMG. As a distinctive structure on RNA, particularly non-coding RNA, we suggest that TMG prevents spurious interactions detrimental to the function of RNAs that it modifies.