Antibody-nucleic acid complexes. Immunospecific retention of globin mRNA with antibodies specific for 7-methylguanosine

Munns, Theodore W.; Liszewski, M. Kathryn; Tellam, Judy; Sims, Harold F.; Rhoads, Robert E.

doi:10.1021/bi00541a018

Cited by 40 publications

(34 citation statements)

References 21 publications

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“…To determine whether or not the cap structure was being removed, we immunoprecipitated the MFA2 transcripts from early and late time points in the transcriptional pulse-chase with antisera directed against the cap structure (Munns et al 1982). As shown in Figure 5, the MFA2 mRNAs from both wild-type and x r n l A cells are precipitable at the beginning of the chase period indicating that they are synthesized with a cap structure (lanes 2,5).…”

Section: Transcripts That Accumulate In X M L H Strains Are Full Lengmentioning

confidence: 99%

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Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript.

Muhlrad¹,

Decker²,

Parker³

1994

Genes Dev.

485

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The first step in the decay of some eukaryotic mRNAs is the shortening of the poly(A) tail. To examine how the transcript body was degraded after deadenylation, we followed the decay of a pulse of newly synthesized MFA2 transcripts while utilizing two strategies to trap intermediates in the degradation pathway. First, we inserted strong RNA secondary structures, which can slow exonucleolytic digestion and thereby trap decay intermediates, into the MFA2 5' UTR. Following deadenylation, fragments of the MFA2 mRNA trimmed from the 5' end to the site of secondary structure accumulated as fulMength mRNA levels decreased. In addition, in cells deleted for the XRN1 gene, which encodes a major 5' to 3' exonuclease in yeast, the MFA2 transcript is deadenylated normally but persists as a fulMength mRNA lacking the 5' cap structure. These results define a mRNA decay pathway in which deadenylation leads to decapping of the mRNA followed by 5' -0 3' exonucleolytic degradation of the transcript body. Because the poly(A) tail and the cap structure are found on essentially all mRNAs, this pathway could be a general mechanism for the decay of many eukaryotic transcripts.

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Section: Transcripts That Accumulate In X M L H Strains Are Full Lengmentioning

confidence: 99%

“…Munns (Munns et al 1982) and graciously provided by E. Lund and J. Dahlberg. Anti-mZG antibodies were prebound at 4°C to protein A-Sepharose CL-4B{Sigma) in IPPH (500 mM NaC1, 10 mM Tris-C1 at pH 7.5, 0.1% NP-40).…”

Section: Mrna Analysismentioning

confidence: 99%

Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript.

Muhlrad¹,

Decker²,

Parker³

1994

Genes Dev.

485

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“…To distinguish between these two possibilities we tested directly whether fuU-length-decapped mRNAs with long poly(A) tails were produced in pabl A xrnlA yeast. This was accomplished by separating capped from decapped transcripts by immunoprecipitation of mRNAs using an antibody directed against the 5' monomethyl cap structure (Munns et al 1982;. In this case, we examined mRNAs from a transcriptional pulse-chase (Decker and Parker 1993), in which transcription from the GALl promoter was rapidly induced and then inhibited.…”

Section: Deletion Of the Xrnl Gene Suppresses A Pablzl And Allows Formentioning

confidence: 99%

“…Munns et al 1982;, and the different fractions were resolved on acrylamide Northern gels. Shown are total (T), pellet (P), and supernatant (S) fractions from each time point; poly(A) tail lengths are depicted to the left of each panel.…”

Section: Ominmentioning

confidence: 99%

Multiple functions for the poly(A)-binding protein in mRNA decapping and deadenylation in yeast.

Caponigro¹,

Parker²

1995

Genes Dev.

259

239

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The first step in the decay of many eukaryotic mRNAs is shortening of the poly(A) tail. In yeast, deadenylation leads to mRNA decapping and subsequent 5' -> 3' exonucleolytic degradation of the transcript body. We have determined that the major poly(A)-binding protein Pablp plays at least two critical roles in this pathway. First, mRNAs in pablA strains were decapped prior to deadenylation. This observation defines a new function for Pablp as an inhibitor of mRNA decapping. Moreover, mutations that inhibit mRNA turnover suppress the inviability of a pablS. mutation, suggesting that premature mRNA decapping in pablA strains contributes to cell death. Second, we find that Pablp is not required for deadenylation, although in its absence poly(A) tail shortening rates are significantly reduced. In addition, in the absence of Pablp, newly synthesized mRNAs had poly(A) tails longer than those in wild-type strains and showed an unexpected temporal delay prior to the initiation of deadenylation and degradation. These results define new and critical functions for Pablp in the regulation of mRNA decapping and deadenylation, two important control points in the specification of mRNA half-lives. Moreover, these results suggest that Pablp functions in additional phases of mRNA metabolism such as mRNP maturation.

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“…However, we found that m and I transcripts produced by the viral RNA polymerase reaction in the presence of 7mGpppG were not immune precipitable by antibody to 7 mG (Munns et al, 1982), unlike transcripts of various recombinant plasmids transcribed in the presence of 7 mGpppG by the RNA polymerases of bacteriophage T7, bacteriophage SP6 or of E. coli.…”

Section: )mentioning

confidence: 62%

Factors Determining Translational Efficiency of mRNA in Yeast,

Leibowitz¹,

Barbone²,

Georgopoulos³

1991

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Antibody-nucleic acid complexes. Immunospecific retention of globin mRNA with antibodies specific for 7-methylguanosine

Cited by 40 publications

References 21 publications

Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript.

Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript.

Multiple functions for the poly(A)-binding protein in mRNA decapping and deadenylation in yeast.

Factors Determining Translational Efficiency of mRNA in Yeast,

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