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. 1992 Nov 25;20(22):5985–5989. doi: 10.1093/nar/20.22.5985

TGA cysteine codons and intron sequences in conserved and nonconserved positions are found in macronuclear RNA polymerase genes of Euplotes octocarinatus.

J Kaufmann 1, V Florian 1, A Klein 1
PMCID: PMC334464  PMID: 1461731

Abstract

The gene sequences of the second largest subunits of RNA polymerases I and II of Euplotes octocarinatus, RPA2 and RPB2, were determined and compared to the respective known sequences of Saccharomyces cerevisiae. The similarity of the derived polypeptide sequences permitted their assignment to the respective polymerases and allowed the comparison of the zinc binding regions. In frame TGA codons were detected, which are likely to encode conserved cysteinyl residues in the putative zinc-finger region of the RPA2 gene. They were also found in other positions in both the RPA2 and RPB2 genes. The RPB2 gene contains a 30 bp intron close to the 5'-end of its coding region. The 5'-ends of the coding regions of all three genes encoding the largest subunits of the three different polymerases were also analyzed. The zinc finger structures again show the use of TGA codons for conserved cysteinyl residues in two of the genes. An N-terminal intron is located in the RPB1 gene at a conserved position as compared to the respective genes of several other eucarya.

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Selected References

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  1. Ahearn J. M., Jr, Bartolomei M. S., West M. L., Cisek L. J., Corden J. L. Cloning and sequence analysis of the mouse genomic locus encoding the largest subunit of RNA polymerase II. J Biol Chem. 1987 Aug 5;262(22):10695–10705. [PubMed] [Google Scholar]
  2. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  3. Bierbaum P., Dönhoff T., Klein A. Macronuclear and micronuclear configurations of a gene encoding the protein synthesis elongation factor EF 1 alpha in Stylonychia lemnae. Mol Microbiol. 1991 Jun;5(6):1567–1575. doi: 10.1111/j.1365-2958.1991.tb00804.x. [DOI] [PubMed] [Google Scholar]
  4. Bird D. M., Riddle D. L. Molecular cloning and sequencing of ama-1, the gene encoding the largest subunit of Caenorhabditis elegans RNA polymerase II. Mol Cell Biol. 1989 Oct;9(10):4119–4130. doi: 10.1128/mcb.9.10.4119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brünen-Nieweler C., Schmidt H. J., Heckmann K. Two introns in the pheromone 3-encoding gene of Euplotes octocarinatus. Gene. 1991 Dec 30;109(2):233–237. doi: 10.1016/0378-1119(91)90613-g. [DOI] [PubMed] [Google Scholar]
  6. Casanova J. L., Pannetier C., Jaulin C., Kourilsky P. Optimal conditions for directly sequencing double-stranded PCR products with sequenase. Nucleic Acids Res. 1990 Jul 11;18(13):4028–4028. doi: 10.1093/nar/18.13.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Harper D. S., Jahn C. L. Differential use of termination codons in ciliated protozoa. Proc Natl Acad Sci U S A. 1989 May;86(9):3252–3256. doi: 10.1073/pnas.86.9.3252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Helftenbein E. Nucleotide sequence of a macronuclear DNA molecule coding for alpha-tubulin from the ciliate Stylonychia lemnae. Special codon usage: TAA is not a translation termination codon. Nucleic Acids Res. 1985 Jan 25;13(2):415–433. doi: 10.1093/nar/13.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Herrick G., Hunter D., Williams K., Kotter K. Alternative processing during development of a macronuclear chromosome family in Oxytricha fallax. Genes Dev. 1987 Dec;1(10):1047–1058. doi: 10.1101/gad.1.10.1047. [DOI] [PubMed] [Google Scholar]
  11. James P., Whelen S., Hall B. D. The RET1 gene of yeast encodes the second-largest subunit of RNA polymerase III. Structural analysis of the wild-type and ret1-1 mutant alleles. J Biol Chem. 1991 Mar 25;266(9):5616–5624. [PubMed] [Google Scholar]
  12. Jokerst R. S., Weeks J. R., Zehring W. A., Greenleaf A. L. Analysis of the gene encoding the largest subunit of RNA polymerase II in Drosophila. Mol Gen Genet. 1989 Jan;215(2):266–275. doi: 10.1007/BF00339727. [DOI] [PubMed] [Google Scholar]
  13. Kontermann R., Sitzler S., Seifarth W., Petersen G., Bautz E. K. Primary structure and functional aspects of the gene coding for the second-largest subunit of RNA polymerase III of Drosophila. Mol Gen Genet. 1989 Nov;219(3):373–380. doi: 10.1007/BF00259609. [DOI] [PubMed] [Google Scholar]
  14. Liebhaber S. A., Emery J. G., Urbanek M., Wang X. K., Cooke N. E. Characterization of a human cDNA encoding a widely expressed and highly conserved cysteine-rich protein with an unusual zinc-finger motif. Nucleic Acids Res. 1990 Jul 11;18(13):3871–3879. doi: 10.1093/nar/18.13.3871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meyer F., Schmidt H. J., Heckmann K. Pheromone 4 gene of Euplotes octocarinatus. Dev Genet. 1992;13(1):16–25. doi: 10.1002/dvg.1020130104. [DOI] [PubMed] [Google Scholar]
  16. Meyer F., Schmidt H. J., Plümper E., Hasilik A., Mersmann G., Meyer H. E., Engström A., Heckmann K. UGA is translated as cysteine in pheromone 3 of Euplotes octocarinatus. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3758–3761. doi: 10.1073/pnas.88.9.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mémet S., Saurin W., Sentenac A. RNA polymerases B and C are more closely related to each other than to RNA polymerase A. J Biol Chem. 1988 Jul 25;263(21):10048–10051. [PubMed] [Google Scholar]
  18. Nawrath C., Schell J., Koncz C. Homologous domains of the largest subunit of eucaryotic RNA polymerase II are conserved in plants. Mol Gen Genet. 1990 Aug;223(1):65–75. doi: 10.1007/BF00315798. [DOI] [PubMed] [Google Scholar]
  19. Pühler G., Leffers H., Gropp F., Palm P., Klenk H. P., Lottspeich F., Garrett R. A., Zillig W. Archaebacterial DNA-dependent RNA polymerases testify to the evolution of the eukaryotic nuclear genome. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4569–4573. doi: 10.1073/pnas.86.12.4569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Seifarth W., Petersen G., Kontermann R., Riva M., Huet J., Bautz E. K. Identification of the genes coding for the second-largest subunits of RNA polymerases I and III of Drosophila melanogaster. Mol Gen Genet. 1991 Sep;228(3):424–432. doi: 10.1007/BF00260636. [DOI] [PubMed] [Google Scholar]
  23. Sitzler S., Oldenburg I., Petersen G., Bautz E. K. Analysis of the promoter region of the housekeeping gene DmRP140 by sequence comparison of Drosophila melanogaster and Drosophila virilis. Gene. 1991 Apr;100:155–162. doi: 10.1016/0378-1119(91)90361-e. [DOI] [PubMed] [Google Scholar]
  24. Swanton M. T., Greslin A. F., Prescott D. M. Arrangement of coding and non-coding sequences in the DNA molecules coding for rRNAs in Oxytricha sp. DNA of ciliated protozoa. VII. Chromosoma. 1980;77(2):203–215. doi: 10.1007/BF00329545. [DOI] [PubMed] [Google Scholar]
  25. Sweetser D., Nonet M., Young R. A. Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1192–1196. doi: 10.1073/pnas.84.5.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vallee B. L., Coleman J. E., Auld D. S. Zinc fingers, zinc clusters, and zinc twists in DNA-binding protein domains. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):999–1003. doi: 10.1073/pnas.88.3.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Woese C. R., Kandler O., Wheelis M. L. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4576–4579. doi: 10.1073/pnas.87.12.4576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yano R., Nomura M. Suppressor analysis of temperature-sensitive mutations of the largest subunit of RNA polymerase I in Saccharomyces cerevisiae: a suppressor gene encodes the second-largest subunit of RNA polymerase I. Mol Cell Biol. 1991 Feb;11(2):754–764. doi: 10.1128/mcb.11.2.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zillig W., Klenk H. P., Palm P., Pühler G., Gropp F., Garrett R. A., Leffers H. The phylogenetic relations of DNA-dependent RNA polymerases of archaebacteria, eukaryotes, and eubacteria. Can J Microbiol. 1989 Jan;35(1):73–80. doi: 10.1139/m89-011. [DOI] [PubMed] [Google Scholar]

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