Quantitative Assessment of mRNA Cap Analogues as Inhibitors of in Vitro Translation

Cai, Ai-Li; Jankowska-Anyszka, Marzena; Centers, Adrian; Chlebicka, Lidia; Stępiński, Janusz; Stolarski, Ryszard; Darżynkiewicz, Edward; Rhoads, Robert E.

doi:10.1021/bi9830213

Cited by 120 publications

(132 citation statements)

References 69 publications

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“…There is some concern that if an RNA has a TMG cap instead of an m7G cap, this could have repercussions for the initiation of translation. The proportion of hypermethylated guanosine-capped RNAs was found to be lower in polysomal RNA than in nonpolysomal RNA, suggesting that TMG RNAs may not be translated efficiently (57,58). How these in vitro findings reflect in vivo translation is not entirely clear, however.…”

Section: Discussionmentioning

confidence: 97%

Trimethylguanosine capping selectively promotes expression of Rev-dependent HIV-1 RNAs

Yedavalli

Jeang

2010

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

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5′-mRNA capping is an early modification that affects pre-mRNA synthesis/splicing, RNA cytoplasmic transport, and mRNA translation and turnover. In eukaryotes, a 7-methylguanosine (m7G) cap is added to newly transcribed RNA polymerase II (RNAP II) transcripts. A subset of RNAP II-transcribed cellular RNAs, including small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), and telomerase RNA, is further hypermethylated at the exocyclic N2 of the guanosine to create a trimethylguanosine (TMG)-capped RNA. Some of these TMG-capped RNAs are transported within the nucleus and from the nucleus to the cytoplasm by the CRM-1 (required for chromosome region maintenance) protein. CRM-1 is also used to export Rev/RRE-dependent unspliced/ partially spliced HIV-1 RNAs. Here we report that like snRNAs and snoRNAs, some Rev/RRE-dependent HIV-1 RNAs are TMG-capped. The methyltransferase responsible for TMG modification of HIV-1 RNAs is the human PIMT (peroxisome proliferator-activated receptor-interacting protein with methyltransferase) protein. TMG capping of unspliced/partially spliced HIV-1 RNAs represents a new regulatory mechanism for selective expression.required for chromosome region maintenance | RNA export | peroxisome proliferator-activated receptor-interacting protein with methyltransferase P osttranscriptional processing of RNA plays a critical role in gene expression (1-3). An early mRNA modification is the formation of a 7-methylguanosine (m7G) cap (4-6). In mammals, the acquisition of an m7G cap requires two enzymes (7-9), a capping enzyme (triphosphatase and guanylyltransferase activity) and an RNA-(guanine 7)-methyltransferase. These proteins are essential for cell growth, and mutations in the triphosphatase, guanylyltransferase, or methyltransferase components of the capping apparatus are lethal in vivo (10).Cellular RNAP II transcripts are mostly m7G-capped (1-3, 11). An m7G cap facilitates the initiation of translation in mammalian cells, and a failure to cap premRNAs results in their accelerated decay by 5′ exoribonuclease degradation (2,(12)(13)(14). Capping of viral RNAs has been less extensively investigated. It has been generally assumed that like cellular RNAs, many viral RNAs have a 5′ m7G cap, and that viruses either use the cellular capping machinery (e.g., viruses that replicate in the nucleus) or encode their own capping enzymes (e.g., viruses that replicate in the cytoplasm) (15, 16). Interestingly, there is a surprisingly diverse range of cap modifications; for instance, Mimivirus, a large DNA virus of amoeba, encodes an RNA cap guanine-N2 methyltransferase that hypermethylates the viral RNA cap (17); influenza virus "snatches" caps from cellular mRNAs (18, 19); West Nile fever virus has two methyl additions on its RNA cap (20); Sindbis virus produces mRNAs that have dimethylguanosine and trimethylguanosine caps [hypermethylated caps/m 2,2,7 G caps (21)]; and Semliki forest virus late mRNAs also have hypermethylated caps (22). On the other hand, poliovirus, encephalomyelitis virus, foot and mouth...

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Section: Discussionmentioning

confidence: 97%

Trimethylguanosine capping selectively promotes expression of Rev-dependent HIV-1 RNAs

Yedavalli

Jeang

2010

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

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“…Unlike eIF4E, N and VP39 bind m 7 GDP or m 7 GMP with a similar weak affinity, suggesting that the triphosphate moiety of the terminal cap does not play a major role during binding. However, the role of triphosphate moiety in the binding of cap with eIF4E is well understood (55)(56)(57). Previous studies have also reported that nucleotides adjacent to the 5Ј cap affect the cap binding with eIF4E, CBP, and VP39 in a different manner.…”

Section: Discussionmentioning

confidence: 99%

Hantavirus Nucleocapsid Protein Has Distinct m7G Cap- and RNA-binding Sites

Mir

Sheema

Haseeb

et al. 2010

Journal of Biological Chemistry

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Hantaviruses, members of the Bunyaviridae family, are emerging category A pathogens that carry three negative stranded RNA molecules as their genome. Hantavirus nucleocapsid protein (N) is encoded by the smallest S segment genomic RNA (viral RNA). N specifically binds mRNA caps and requires four nucleotides adjacent to the cap for high affinity binding. We show that the N peptide has distinct cap-and RNA-binding sites that independently interact with mRNA cap and viral genomic RNA, respectively. In addition, N can simultaneously bind with both mRNA cap and vRNA. N undergoes distinct conformational changes after binding with either mRNA cap or vRNA or both mRNA cap and vRNA simultaneously. Hantavirus RNA-dependent RNA polymerase (RdRp) uses a capped RNA primer for transcription initiation. The capped RNA primer is generated from host cell mRNA by the cap-snatching mechanism and is supposed to anneal with the 3 terminus of vRNA template during transcription initiation by single G-C base pairing. We show that the capped RNA primer binds at the cap-binding site and induces a conformational change in N. The conformationally altered N with a capped primer loaded at the cap-binding site specifically binds the conserved 3 nine nucleotides of vRNA and assists the bound primer to anneal at the 3 terminus. We suggest that the cap-binding site of N, in conjunction with RdRp, plays a key role during the transcription and replication initiation of vRNA genome.Hantaviruses cause two types of serious human illnesses when transmitted to humans from rodent hosts: hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome (1, 2). The spherical hantavirus particles harbor three negative sense genomic RNA segments (S, L, and M segments) within a lipid bilayer (3). The mRNAs derived from S, L, and M segments encode viral nucleocapsid protein (N), viral RNA-dependent RNA polymerase (RdRp), 2 and glycoproteins (G1 and G2), respectively. The characteristic feature of the hantaviral genome is the partially complementary sequence at the 5Ј and 3Ј termini of each of the three genome segments that undergo base pairing and form panhandle structures (4 -6). N is a multifunctional protein playing a vital role in multiple processes of virus replication cycle and has been found to undergo trimerization both in vivo and in vitro (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19). During encapsidation, the three viral RNA (vRNA) molecules are specifically recognized by N inside the host cell and targeted for packaging. Multiple in vitro studies have shown that N preferentially binds vRNA compared with complementary RNA (cRNA) or nonviral RNA (13, 20 -25). It has been proposed that the specific binding of N with either the panhandle or the sequence at the 5Ј terminus alone selectively facilitates the encapsidation of vRNA to generate three nucleocapsids that are packaged into infectious virions (25, 26). The RNA-binding domain of Hantaan virus N protein has been mapped to the central conserved region corresponding to amino acids f...

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“…Consistent with this explanation, the GpppG analogue, which is unable to bind eIF4E, and cannot induce conformational changes, does not alter eIF4E nuclear body morphology. 14,85,88 The GpppG analogue is also unable to alter PML-NB morphology. However, unlike eIF4E, PML protein is unable to directly bind the m 7 GpppG cap analogue.…”

Section: Spotlightmentioning

confidence: 99%

Finding a role for PML in APL pathogenesis: a critical assessment of potential PML activities

Strudwick

Borden

2002

Leukemia

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In normal mammalian cells the promyelocytic leukemia protein (PML) is primarily localized in multiprotein nuclear complexes called PML nuclear bodies. However, both PML and PML nuclear bodies are disrupted in acute promyelocytic leukemia (APL). The treatment of APL patients with all-trans retinoic acid (ATRA) results in clinical remission associated with blast cell differentiation and reformation of the PML nuclear bodies. These observations imply that the structural integrity of the PML nuclear body is critically important for normal cellular functions. Indeed, PML protein is a negative growth regulator capable of causing growth arrest in the G 1 phase of the cell cycle, transformation suppression, senescence and apoptosis. These PML-mediated, physiological effects can be readily demonstrated. However, a discrete biochemical and molecular model of PML function has yet to be defined. Upon first assessment of the current PML literature there appears to be a seemingly endless list of potential PML partner proteins implicating PML in a variety of regulatory mechanisms at every level of gene expression. The purpose of this review is to simplify this confusing field of research by using strict criteria to deduce which models of PML body function are well supported.

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Quantitative Assessment of mRNA Cap Analogues as Inhibitors of in Vitro Translation

Cited by 120 publications

References 69 publications

Trimethylguanosine capping selectively promotes expression of Rev-dependent HIV-1 RNAs

Trimethylguanosine capping selectively promotes expression of Rev-dependent HIV-1 RNAs

Hantavirus Nucleocapsid Protein Has Distinct m7G Cap- and RNA-binding Sites

Finding a role for PML in APL pathogenesis: a critical assessment of potential PML activities

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