A relation between inhibition of protein synthesis and conformation of 5′-phosphorylated 7-methylguanosine derivatives

Hickey, Eileen; Weber, Lee A.; Baglioni, Corrado; Kim, Chong H.; Sarma, Ramaswamy H.

doi:10.1016/s0022-2836(77)80027-2

Cited by 70 publications

(36 citation statements)

References 25 publications

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“…The degree of inhibition is proportional to the number of phosphate groups in the methylated cap analogues, which is characteristic also of the inhibitory effect of cap analogues on translation (14). However, as pointed out before (6), the concentration of cap analogue required to inhibit splicing is two orders of magnitude lower than that required to inhibit mRNA translation to the same extent.…”

Section: Discussionmentioning

confidence: 84%

Cap-dependent RNA splicing in a HeLa nuclear extract.

Edery¹,

Sonenberg²

1985

Proc. Natl. Acad. Sci. U.S.A.

144

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We have studied the involvement of the 5' cap structure in the splicing of precursor mRNAs in a HeLa nuclear extract. We show that precursor mRNAs are spliced efficiently only when they possess a cap structure and that preincubation of a HeLa nuclear extract rendered the splicing reaction highly sensitive to inhibition by cap analogues. This sensitization was dependent on exogenous Mg2' but not exogenous ATP or GTP.These results demonstrate that splicing in a nuclear extract is highly dependent on the cap structure as was demonstrated for the splicing process in a HeLa whole-cell extract [Konarska, M. M., Padget, R. A. & Sharp, P. A. (1984) Cell 38,[731][732][733][734][735][736], and thus support the contention that cap recognition is an important feature of eukaryotic mRNA biogenesis.All eukaryotic cellular mRNAs analyzed to date are blocked at their 5' terminus by the cap structure, m7G(5')ppp(5')N (1). There is considerable evidence that the cap structure enhances translational efficiency by facilitating ribosome binding to mRNA (1). This interaction is mediated by a distinct group of proteins defined as cap-binding proteins (2,3).With the advent of efficient in vitro splicing systems (4-10), two recent studies have suggested that the cap structure may play a significant role in the splicing process as well (6, 7). Using a HeLa nuclear extract, Krainer et al. (7) were able to show efficient splicing of in vitro synthesized, truncated human P-globin transcripts. They showed that although uncapped transcripts could still be spliced, the efficiency was one-half to one-third that for their enzymatically capped counterparts. In addition, splicing of uncapped precursor mRNAs (pre-mRNAs) The transcripts were stored at -70'C in water and used directly in the splicing reaction.In Vitro Splicing Reaction. HeLa nuclear extracts were prepared as described (13). All experiments described throughout were performed using the same extract prepara- RESULTSTo characterize the splicing activity of our HeLa nuclear extracts, we used the plasmid pSP64-il/3A6. This plasmid and its use in the nuclear splicing system were described by Krainer et al. (7) and Ruskin et al. (8). For preparation of 7590The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Discussionmentioning

confidence: 84%

Cap-dependent RNA splicing in a HeLa nuclear extract.

Edery¹,

Sonenberg²

1985

Proc. Natl. Acad. Sci. U.S.A.

144

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“…On the basis of NMR spectral analyses of 5 ' -phosphorylated 7-methylguanosine derivatives, Hickey et al [8] reported that the introduction of N7-methyl and 5 '-phosphate into guanosine induces the nucleotide to assume a rigid conformation in which the sugar moiety is preferentially C3 ' -endo with + SC (gauche. gauche) orientation around the exocyclic C4'-C5' bond, and this conformation may be essential for inhibition of pro- Table 1 Conformational angles (") of the m'GMP molecule tein synthesis as result of the conformational similarity with the 5 ' -terminal capped structure of mRNA.…”

Section: Resultsmentioning

confidence: 99%

Indole ring binds to 7‐methylguanine base by π‐π stacking interaction

et al. 1986

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“…Table 2 summarizes conformational parameters of 7-methylguanosine nucleotides in solution. Unlike other purine nucleosides and nucleotides, 7-methylguanosine and its 5´-phosphorylated derivatives prefer a 3´-endo sugar ring conformation [26][27][28][29][30][31]. With 7-methylguanosine, the proportion of the 3´-endo conformers is 50 %, whereas guanosine preferably exists in a 2´-endo conformation (38 % 3´-endo).…”

Section: Conformation and Intramolecular Interactionsmentioning

confidence: 99%

Preparation and Properties of mRNA 5-cap Structure

Mikkola¹,

Salomaki²,

Zhang³

et al. 2005

COC

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The 5´-terminus of eukaryotic messenger RNA (mRNA) molecules contains an unique structure, viz. a dinucleoside 5´,5´-triphosphate moiety where the terminal nucleoside is 7-methylguanosine. This 5´-cap structure serves as a site of recognition for numerous enzymes involved in splicing, transport and translation of mRNA, and protects mRNA against intracellular exonucleases. In addition, viral RNA polymerases use capped mRNA sequences of the host cell as primers for viral RNA synthesis. Understanding of molecular mechanism of all these processes requires detailed information on the chemical properties of the cap structure, including capability to prepare conjugates and structural analogs of the cap for research tools. This review tends to summarize the present knowledge on various aspects of the chemistry of the cap structure, including chemical stability and mechanisms of breakdown, protolytic and complexing equilibria, stacking interactions and synthetic methodology.

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A relation between inhibition of protein synthesis and conformation of 5′-phosphorylated 7-methylguanosine derivatives

Cited by 70 publications

References 25 publications

Cap-dependent RNA splicing in a HeLa nuclear extract.

Cap-dependent RNA splicing in a HeLa nuclear extract.

Indole ring binds to 7‐methylguanine base by π‐π stacking interaction

Preparation and Properties of mRNA 5-cap Structure

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