Nonhydrolyzable ATP Analog 5′-Adenylyl-imidodiphosphate (AMP-PNP) Does Not Inhibit ATP-Dependent Scanning of Leader Sequence of mRNA

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P. A. Sakharov, A. S. Sokolov, and S. Ch. Agalarov^*

Institute of Protein Research, Russian Academy of Sciences, ul. Institutskaya 4, 142290 Pushchino, Moscow Region, Russia; fax: +7 (495) 514-0218; E-mail: sultan@vega.protres.ru

^* To whom correspondence should be addressed.

Received June 14, 2014; Revision received July 22, 2014

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The objective of the present work was to determine whether it is possible to use a nonhydrolyzable analog of ATP (AMP-PNP) as an inhibitor of ATP-dependent scanning of the leader sequence of eukaryotic mRNA in translation initiation­. The formation of ribosomal 48S initiation complexes at the start codon of the capped mRNA leader sequence of rabbit β-globin mRNA was studied. The study was carried out in a system composed of individual components of translation initiation. The dependences of the efficiency of formation of 48S initiation complexes on ATP concentration and incubation time were obtained in the absence and presence of AMP-PNP. It was found that AMP-PNP did not affect the efficiency of formation of 48S initiation complexes in all cases under study. We conclude that the uncleavable analog of ATP, AMP-PNP, is not an inhibitor of translation initiation in eukaryotes.

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KEY WORDS: translation initiation, 48S ribosomal initiation complex, AMP-PNP, toeprinting

DOI: 10.1134/S0006297915010058

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