Accurate Transcription of Truncated Ribosomal DNA Templates in a Drosophila Cell-free System

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Mar 1

PMID 6803243

Citations 44

Authors

B D Kohorn

P M Rae

Affiliations

Soon will be listed here.

Abstract

An extract of Drosophila melanogaster Kc cells is shown to give specific and accurate transcription of truncated segments of cloned D. melanogaster ribosomal DNA (rDNA). When clones are digested with restriction enzymes so that the initiation site is flanked by 0.3 kilobase (kb) of nontranscribed spacer and greater than 0.4 kb of external transcribed spacer, RNA polymerase I activity in the extract parallels in vivo rRNA synthesis in selection of the coding strand of template and the site of transcription initiation. When greater than 0.3 kb of the nontranscribed spacer is contiguous with transcribed spacer, in vitro initiations evidently also occur in repeated sequences adjacent to the site of in vivo initiation; when less than or equal to 0.4 kb of the external transcribed spacer is present in a segment, expected transcripts are heterogeneous in length or not detectable. Transcription in the cell-free system requires the specific addition of D. melanogaster rDNA: neither D. virilis rDNA, vector plasmid, nor clones of D. melanogaster genes that are transcribed in vivo by RNA polymerases II and III serve as templates in the system. Drosophila rDNA units that have an interruption in the 28S rRNA coding region are not transcribed in vivo, but restriction digests of a recombinant phage DNA that contains such a unit are active as template for in vitro rDNA transcription.

Citing Articles

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Widely differing degrees of sequence conservation of the two types of rDNA insertion within the melanogaster species sub-group of Drosophila.

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References

ECHALIER G, Ohanessian A . In vitro culture of Drosophila melanogaster embryonic cells. In Vitro. 1970; 6(3):162-72. DOI: 10.1007/BF02617759. View

Blumenthal T, Landers T . The inhibition of nucleic acid-binding proteins by aurintricarboxylic acid. Biochem Biophys Res Commun. 1973; 55(3):680-8. DOI: 10.1016/0006-291x(73)91198-4. View

Scheer U, Trendelenburg M, Krohne G, Franke W . Lengths and patterns of transcriptional units in the amplified nucleoli of oocytes of Xenopus laevis. Chromosoma. 1977; 60(2):147-67. DOI: 10.1007/BF00288462. View

Valenzuela P, Bell G, Venegas A, Sewell E, Masiarz F, DeGennaro L . Ribosomal RNA genes of Saccharomyces cerevisiae. II. Physical map and nucleotide sequence of the 5 S ribosomal RNA gene and adjacent intergenic regions. J Biol Chem. 1977; 252(22):8126-35. View

Wu G . Adenovirus DNA-directed transcription of 5.5S RNA in vitro. Proc Natl Acad Sci U S A. 1978; 75(5):2175-9. PMC: 392514. DOI: 10.1073/pnas.75.5.2175. View

Dawid I, Wellauer P, Long E . Ribosomal DNA in Drosophila melanogaster. I. Isolation and characterization of cloned fragments. J Mol Biol. 1978; 126(4):749-68. DOI: 10.1016/0022-2836(78)90018-9. View

Weil P, Segall J, Harris B, Ng S, Roeder R . Faithful transcription of eukaryotic genes by RNA polymerase III in systems reconstituted with purified DNA templates. J Biol Chem. 1979; 254(13):6163-73. View

Boseley P, Moss T, Machler M, Portmann R, BIRNSTIEL M . Sequence organization of the spacer DNA in a ribosomal gene unit of Xenopus laevis. Cell. 1979; 17(1):19-31. DOI: 10.1016/0092-8674(79)90291-5. View

Telford J, Kressmann A, Koski R, Grosschedl R, Muller F, Clarkson S . Delimitation of a promoter for RNA polymerase III by means of a functional test. Proc Natl Acad Sci U S A. 1979; 76(6):2590-4. PMC: 383653. DOI: 10.1073/pnas.76.6.2590. View

10.

Silverman S, Schmidt O, Soll D, Hovemann B . The nucleotide sequence of a cloned Drosophila arginine tRNA gene and its in vitro transcription in Xenopus germinal vesicle extracts. J Biol Chem. 1979; 254(20):10290-4. View

11.

Long E, Dawid I . Restriction analysis of spacers in ribosomal DNA of Drosophila melanogaster. Nucleic Acids Res. 1979; 7(1):205-15. PMC: 328006. DOI: 10.1093/nar/7.1.205. View

12.

Weil P, Luse D, Segall J, Roeder R . Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979; 18(2):469-84. DOI: 10.1016/0092-8674(79)90065-5. View

13.

Reeder R . The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell. 1979; 18(2):485-99. DOI: 10.1016/0092-8674(79)90066-7. View

14.

Long E, Dawid I . Expression of ribosomal DNA insertions in Drosophila melanogaster. Cell. 1979; 18(4):1185-96. DOI: 10.1016/0092-8674(79)90231-9. View

15.

Jolly D, Thomas Jr C . Nuclear RNA transcripts from Drosophila melanogaster ribosomal RNA genes containing introns. Nucleic Acids Res. 1980; 8(1):67-84. PMC: 327243. DOI: 10.1093/nar/8.1.67. View

16.

DE Wet J, Daniels D, Schroeder J, Williams B, Moore D, Blattner F . Restriction maps for twenty-one Charon vector phages. J Virol. 1980; 33(1):401-10. PMC: 288556. DOI: 10.1128/JVI.33.1.401-410.1980. View

17.

Maxam A, Gilbert W . Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980; 65(1):499-560. DOI: 10.1016/s0076-6879(80)65059-9. View

18.

Long E, Dawid I . Repeated genes in eukaryotes. Annu Rev Biochem. 1980; 49:727-64. DOI: 10.1146/annurev.bi.49.070180.003455. View

19.

Bittner M, Kupferer P, Morris C . Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Anal Biochem. 1980; 102(2):459-71. DOI: 10.1016/0003-2697(80)90182-7. View

20.

Klemenz R, Geiduschek E . The 5' terminus of the precursor ribosomal RNA of Saccharomyces cerevisiae. Nucleic Acids Res. 1980; 8(12):2679-89. PMC: 324113. DOI: 10.1093/nar/8.12.2679. View