Cap analogs containing 6-thioguanosine--reagents for the synthesis of mRNAs selectively photo-crosslinkable with cap-binding biomolecules.
Numerous biomolecules recognize the 7-methylguanosine cap structure present at the 5' ends of eukaryotic mRNAs. Photo-crosslinking is a valuable technique to study these interactions. We report three anti-reverse cap analogs containing a photo-activable nucleoside, 6-thioguanosine ((6S)G), that enable the synthesis of capped RNAs with (6S)G positioned exclusively as the first transcribed nucleotide. The effect of the 6-thioguanosine moiety on binding to the translation factor eIF4E and the efficiency of mRNA translation was determined. The utility of mRNAs with a (6S)G-modified cap in crosslinking experiments is shown by mapping the histone H4 cap-binding pocket.
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Monika
M
Nowakowska
Joanna
J
Kowalska
Franck
F
Martin
Arnaud
A
d'Orchymont
Joanna
J
Zuberek
Maciej
M
Lukaszewicz
Edward
E
Darzynkiewicz
Jacek
J
Jemielity
D003432:Cross-Linking Reagents / Q000737:chemistry*; D006151:Guanosine / Q000031:analogs & derivatives* / Q000737:chemistry; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000138:chemical synthesis*; D012333:RNA, Messenger / Q000737:chemistry*; D013872:Thionucleosides / Q000737:chemistry*; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D012314:RNA Cap Analogs; D012333:RNA, Messenger; D013872:Thionucleosides; D006151:Guanosine; C005783:6-thioguanosine