Synthesis and properties of mRNA cap analogs containing imidodiphosphate moiety--fairly mimicking natural cap structure, yet resistant to enzymatic hydrolysis.
We describe synthesis and properties of eight dinucleotide mRNA 5' cap analogs containing imidodiphosphate moiety within 5',5'-tri- or tetraphosphate bridge (NH-analogs). The compounds were obtained by coupling an appropriate nucleoside 5'-imidodiphosphate with nucleotide P-imidazolide mediated by divalent metal chloride in anhydrous DMF. To evaluate the novel compounds as tools for studying cap-dependent processes, we determined their binding affinities for eukaryotic translation initiation factor 4E, susceptibilities to decapping pyrophosphatase DcpS and, for non-hydrolysable analogs, binding affinities to this enzyme. The results indicate that the O to NH substitution in selected positions of oligophosphate bridge ensures resistance to enzymatic decapping and suggest that interactions of NH-analogs with cap binding proteins fairly mimic interactions of unmodified parent compounds. Finally, we identified NH-analogs as potent inhibitors of cap-dependent translation in cell free system, and evaluated their utility as reagents for obtaining 5' capped mRNAs in vitro to be rather moderate.
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Zwirki i Wigury 93, Warsaw, Poland.
Anna M
AM
Rydzik
Marta
M
Kulis
Maciej
M
Lukaszewicz
Joanna
J
Kowalska
Joanna
J
Zuberek
Zbigniew M
ZM
Darzynkiewicz
Edward
E
Darzynkiewicz
Jacek
J
Jemielity
D040761:Biomimetic Materials / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D004164:Diphosphonates / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D011485:Protein Binding; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism
D004164:Diphosphonates; D012314:RNA Cap Analogs; C016514:imidodiphosphonic acid