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Cryo-EM structure of a Group II Intron Complexed with its Reverse TranscriptaseCryo-EM structure of a Group II Intron Complexed with its Reverse Transcriptase
Structural highlights
FunctionLTRA_LACLC Multifunctional protein that promotes group II intron splicing and mobility by acting both on RNA and DNA. It has three activities: reverse transcriptase (RT) for intron duplication, maturase to promote splicing, and DNA endonuclease for site-specific cleavage of recipient alleles. The intron-encoded protein promotes splicing by facilitating the formation of the catalytically active structure of the intron RNA. After splicing, the protein remains bound to the excised intron lariat RNA, forming ribonucleoprotein particles, and cleaving the antisense strand of the recipient DNA in the 3' exon. After DNA cleavage, retrohoming occurs by a target DNA-primed reverse transcription of the intron RNA that had reverse spliced into the sense strand of the recipient DNA. It also contributes to the recognition of the DNA target site and acts as a repressor of its own translation. Publication Abstract from PubMedA major hurdle for single particle cryo-EM in structural determination lies in the specimen preparation impaired by the air-water interface (AWI) and preferential particle-orientation problems. In this work, we develop functionalized graphene grids with various charges via a dediazoniation reaction for cryo-EM specimen preparation. The graphene grids are paraffin-assistant fabricated, which appear with less contaminations compared with those produced by polymer transfer method. By applying onto three different types of macromolecules, we demonstrate that the high-yield charged graphene grids bring macromolecules away from the AWI and enable adjustable particle-orientation distribution for more robust single particle cryo-EM structural determination. Functionalized graphene grids with various charges for single-particle cryo-EM.,Lu Y, Liu N, Liu Y, Zheng L, Yang J, Wang J, Jia X, Zi Q, Peng H, Rao Y, Wang HW Nat Commun. 2022 Nov 7;13(1):6718. doi: 10.1038/s41467-022-34579-w. PMID:36344519[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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