Post-transcriptional Adenylation of Signal Recognition Particle RNA Is Carried Out by an Enzyme Different from mRNA Poly(A) Polymerase*

A fraction of the signal recognition particle (SRP) RNA from human, rat, Xenopus, and Saccharomyces cerevisiae cells contains a single post-transcriptionally added adenylic acid residue on its 3′-end; in the case of human SRP RNA, over 60% of the SRP RNA molecules contain a nontemplated adenylic acid residue on their 3′-ends (Sinha, K. M., Gu, J., Chen, Y., and Reddy, R. (1998) J. Biol. Chem. 273, 6853–6859). In this study, we investigated the enzyme that is involved in this 3′-end adenylation of SRP RNA. A U1A protein peptide conjugated to albumin completely inhibited the polyadenylation of a SV40 mRNA by HeLa cell nuclear extract in vitro; however, the 3′-end adenylation of human SRP RNA or Alu RNA, which corresponds to 5′ and 3′-ends of SRP RNA, was not affected by this U1A peptide conjugate. SRP RNA from mutant strains of S. cerevisiaewith a temperature-sensitive mRNA poly(A) polymerase grown at a restrictive temperature of 37 °C also contained a post-transcriptionally added adenylic acid residue just like SRP RNA from wild-type cells and mutant cells grown at permissive temperature of 23 °C. In addition, binding of SRP 9/14-kDa protein heterodimer was required for adenylation of Alu RNA in vitro. These lines of evidence, along with other data, show that post-transcriptional adenylation of SRP and Alu RNAs is carried out by a novel enzyme that is distinct from the mRNA poly(A) polymerase, CCA-adding enzyme, and nonspecific terminal transferase.

[1]  Yahua Chen,et al.  Accurate 3′ End Processing and Adenylation of Human Signal Recognition Particle RNA and Alu RNA in Vitro * , 1998, The Journal of Biological Chemistry.

[2]  A. Weiner,et al.  The CCA-adding Enzyme Has a Single Active Site* , 1998, The Journal of Biological Chemistry.

[3]  M. Deutscher,et al.  Polyadenylation of stable RNA precursors in vivo. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Węgrzyn,et al.  Rapid degradation of polyadenylated oop RNA , 1998, FEBS letters.

[5]  M. Ares,et al.  Depletion of yeast RNase III blocks correct U2 3′ end formation and results in polyadenylated but functional U2 snRNA , 1998, The EMBO journal.

[6]  A. Weiner,et al.  CCA addition by tRNA nucleotidyltransferase: polymerization without translocation? , 1998, The EMBO journal.

[7]  Y. Chen,et al.  Adenylation of Small RNAs in Human Cells , 1998, The Journal of Biological Chemistry.

[8]  M. Deutscher,et al.  Functional Overlap of tRNA Nucleotidyltransferase, Poly(A) Polymerase I, and Polynucleotide Phosphorylase* , 1997, The Journal of Biological Chemistry.

[9]  T. Cech,et al.  Polyadenylation of telomerase RNA in budding yeast. , 1997, RNA.

[10]  S. Cusack,et al.  Identification of a minimal Alu RNA folding domain that specifically binds SRP9/14. , 1997, RNA.

[11]  S. Cusack,et al.  The crystal structure of the signal recognition particle Alu RNA binding heterodimer, SRP9/14 , 1997, The EMBO journal.

[12]  I. Mattaj,et al.  Involvement of the carboxyl terminus of vertebrate poly(A) polymerase in U1A autoregulation and in the coupling of splicing and polyadenylation. , 1997, Genes & development.

[13]  S. Pääbo,et al.  Polyadenylation creates the discriminator nucleotide of chicken mitochondrial tRNA(Tyr). , 1997, Journal of molecular biology.

[14]  A. Weiner,et al.  CCA-adding enzymes and poly(A) polymerases are all members of the same nucleotidyltransferase superfamily: characterization of the CCA-adding enzyme from the archaeal hyperthermophile Sulfolobus shibatae. , 1996, RNA.

[15]  Chris Sander,et al.  DNA polymerase β belongs to an ancient nucleotidyltransferase superfamily , 1995 .

[16]  W. Keller,et al.  No end yet to messenger RNA 3′ processing! , 1995, Cell.

[17]  L. Minvielle-Sebastia,et al.  The FIP1 gene encodes a component of a yeast pre-mRNA polyadenylation factor that directly interacts with poly(A) polymerase , 1995, Cell.

[18]  J. Manley,et al.  A complex protein assembly catalyzes polyadenylation of mRNA precursors. , 1995, Current opinion in genetics & development.

[19]  Stanley N Cohen,et al.  RNA degradation in Escherichia coli regulated by 3' adenylation and 5' phosphorylation , 1995, Nature.

[20]  L. Minvielle-Sebastia,et al.  RNA14 and RNA15 proteins as components of a yeast pre-mRNA 3'-end processing factor. , 1994, Science.

[21]  H. Fried,et al.  Nuclear export of signal recognition particle RNA is a facilitated process that involves the Alu sequence domain. , 1994, Journal of cell science.

[22]  A. Virtanen,et al.  Multiple forms of poly(A) polymerases in human cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  C. Moore,et al.  Separation of factors required for cleavage and polyadenylation of yeast pre-mRNA , 1992, Molecular and cellular biology.

[24]  J. Butler,et al.  Conditional defect in mRNA 3' end processing caused by a mutation in the gene for poly(A) polymerase , 1992, Molecular and cellular biology.

[25]  J. Lingner,et al.  Cloning and expression of the essential gene for poly(A) polymerase from S. cerevisiae , 1991, Nature.

[26]  P. Walter,et al.  Binding sites of the 9- and 14-kilodalton heterodimeric protein subunit of the signal recognition particle (SRP) are contained exclusively in the Alu domain of SRP RNA and contain a sequence motif that is conserved in evolution. , 1991, Molecular and cellular biology.

[27]  N. Larsen,et al.  SRP-RNA sequence alignment and secondary structure. , 1991, Nucleic acids research.

[28]  J. Manley,et al.  Multiple forms of poly(A) polymerases purified from HeLa cells function in specific mRNA 3'-end formation , 1989, Molecular and cellular biology.

[29]  G. Buck,et al.  In vitro 3' end processing and poly(A) tailing of RNA in Trypanosoma cruzi. , 1989, Nucleic acids research.

[30]  J. Manley,et al.  Separation and characterization of a poly(A) polymerase and a cleavage/specificity factor required for pre-mRNA polyadenylation , 1988, Cell.

[31]  M. Wickens,et al.  Products of in vitro cleavage and polyadenylation of simian virus 40 late pre-mRNAs , 1987, Molecular and cellular biology.

[32]  R. Roeder,et al.  Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. , 1983, Nucleic acids research.

[33]  P. Piper,et al.  Altered maturation of sequences at the 3′ terminus of 5S gene transcripts in a Saccharomyces cerevisiae mutant that lacks a RNA processing endonuclease. , 1983, The EMBO journal.

[34]  D. Clayton.  Transcription and replication of animal mitochondrial DNAs. , 1992, International review of cytology.

[35]  M. Deutscher.  Ribonucleases, tRNA nucleotidyltransferase, and the 3' processing of tRNA. , 1990, Progress in nucleic acid research and molecular biology.

[36]  Zieve Gw,et al.  Cell biology of the snRNP particles. , 1990 .

[37]  S. Seifter,et al.  [22] Precipitation techniques , 1990 .

[38]  K. Takeshita,et al.  Polyadenylation of a human mitochondrial ribosomal RNA transcript detected by molecular cloning. , 1985, Gene.

[39]  G. Attardi.  Animal mitochondrial DNA: an extreme example of genetic economy. , 1985, International review of cytology.