Project Details
Projekt
Investigation of RNA-specific base modification pathways
Applicant
Professor Dr. Gunter Meister
Subject Area
Biochemistry
Structural Biology
Structural Biology
Term
from 2015 to 2023
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 277455384
Chemical modifications on RNA bases are not only frequent in well-studied non-coding RNAs such as transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs) but also found in mRNAs. The most prominent modification on mRNAs is m6A, which is found at various positions and can have effects on gene expression. In addition, smaller amounts of m1A, m5C or pseudoU have been identified on mRNAs as well. The identification, however, is often difficult since specific antibodies are only rarely available. Therefore, it is one of the goals of our project to generate highly specific monoclonal antibodies against modified nucleotides. During the first funding period, we have established antibodies against m6A, m26A, m1A and m5C. Based on the experience we have gained, we will now continue to generate antibodies against m3C, m4C, m1G, m2G, m5U, m1pseudoU and m3U. Furthermore, we will establish antibodies against nicotinamide-adenine-dinucleotide (NAD), which has recently been shown to serve as 5’ mRNA cap in prokaryotes and eukaryotes. Base-methylations are catalyzed by specific methyltransferases. A dimer composed of METTL3/14, for example, generates m6A on distinct target sequences. During the first funding period, we predicted that the uncharacterized methyltransferases METTL2, 4, 6 and 8 might function on RNAs as well. In preliminary studies we indeed found that METTL8 is associated with RNA. We will now characterize these enzymes in more detail. We will (i) investigate which modification is catalyzed, (ii) which RNAs are modified and (iii) which RNA motifs are used by these enzymes. During the first funding period, we already succeeded to generate recombinant METTL enzymes. These proteins will now be used for in vitro methylation assays as well structural studies.
DFG Programme
Priority Programmes
