title: Modifying the 5'-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells.
abstract: The 5'-cap is a hallmark of eukaryotic mRNAs and plays fundamental roles in RNA metabolism, ranging from quality control to export and translation. Modifying the 5'-cap may thus enable modulation of the underlying processes and investigation or tuning of several biological functions. A straightforward approach is presented for the efficient production of a range of N7-modified caps based on the highly promiscuous methyltransferase Ecm1. We show that these, as well as N(2) -modified 5'-caps, can be used to tune translation of the respective mRNAs both in vitro and in cells. Appropriate modifications allow subsequent bioorthogonal chemistry, as demonstrated by intracellular live-cell labeling of a target mRNA. The efficient and versatile N7 manipulation of the mRNA cap makes mRNAs amenable to both modulation of their biological function and intracellular labeling, and represents a valuable addition to the chemical biology toolbox.
authors: University of Muenster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Strasse 2, 48149, Muenster, Germany.
authors: Josephin M
authors: JM
authors: Holstein
authors: University of Muenster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Strasse 2, 48149, Muenster, Germany.
authors: Lea
authors: L
authors: Anhäuser
authors: University of Muenster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Strasse 2, 48149, Muenster, Germany. a.rentmeister@uni-muenster.de.|Cells-in-Motion Cluster of Excellence (EXC 1003-CiM), University of Muenster, Germany. a.rentmeister@uni-muenster.de.
authors: Andrea
authors: A
authors: Rentmeister
mesh_terms: D057930:Click Chemistry; D009670:Encephalitozoon cuniculi / Q000201:enzymology; D056890:Eukaryota / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008780:Methyltransferases / Q000378:metabolism; D018613:Microscopy, Confocal; D012315:RNA Caps / Q000737:chemistry* / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives
chemical_list: D005656:Fungal Proteins; D012315:RNA Caps; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012436:S-Adenosylmethionine; D008780:Methyltransferases
keywords: Ecm1; RNA labeling; bioorthogonal chemistry; mRNA; translation