Phosphate-modified analogues of m(7)GTP and m(7)Gppppm(7)G-Synthesis and biochemical properties.
The synthesis and biochemical properties of 17 new mRNA cap analogues are reported. Six of these nucleotides are m(7)GTP derivatives, whereas 11 are 'two headed' tetraphosphate dinucleotides based on a m(7)Gppppm(7)G structure. The compounds contain either a boranophosphate or phosphorothioate moiety in the nucleoside neighbouring position(s) and some of them possess an additional methylene group between β and γ phosphorus atoms. The compounds were prepared by divalent metal chloride-mediated coupling of an appropriate m(7)GMP analogue with a given P(1),P(2)-di(1-imidazolyl) derivative. The analogues were evaluated as tools for studying cap-dependent processes in a number of biochemical assays, including determination of affinity to eukaryotic initiation factor eIF4E, susceptibility to enzymatic hydrolysis, and translational efficiency in vitro. The results indicate that modification in the phosphate chain can increase binding to cap-interacting proteins and provides higher resistance to degradation. Furthermore, modified derivatives of m(7)GTP were found to be potent inhibitors of cap-dependent translation in cell free systems.
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Marcin
M
Ziemniak
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Joanna
J
Kowalska
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Maciej
M
Lukaszewicz
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Joanna
J
Zuberek
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland.
Katarzyna
K
Wnek
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland; Centre of New Technologies, University of Warsaw, Banacha 2c, 02-097 Warsaw, Poland.
Edward
E
Darzynkiewicz
Centre of New Technologies, University of Warsaw, Banacha 2c, 02-097 Warsaw, Poland. Electronic address: jacekj@biogeo.uw.edu.pl.
Jacek
J
Jemielity
D001880:Boranes / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D010710:Phosphates / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D054735:Phosphorothioate Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D012333:RNA, Messenger / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism
D001880:Boranes; D039561:Eukaryotic Initiation Factor-4E; D010710:Phosphates; D054735:Phosphorothioate Oligonucleotides; D012314:RNA Cap Analogs; D012333:RNA, Messenger; C510710:boranophosphate; C017516:7-methylguanosine triphosphate
Bioorganic chemistry; Boranophosphates; Nucleotides; Phosphorothioates; mRNA cap