Transcription of T-DNA in octopine and nopaline crown gall tumours is inhibited by low concentrations of alpha-amanitin.

Nucleic acids research ·Vol. 9 ·No. 19 ·1981-10-10 ·Pages 4801-12

Willmitzer L, Schmalenbach W, Schell J

Abstract

Highly purified and physiologically active nuclei were isolated from four different octopine and nopaline crown gall lines. These nuclei exhibited a high endogenous RNA synthesizing activity involving all three RNA-polymerases I, II and III. Isolated nuclei were shown by Southern blotting to synthesize T-DNA specific RNA. This synthesis was shown to be sensitive to actinomycin D and therefore to be DNA-dependent. The transcription of the T-DNA was also inhibited for more than 90% by low concentrations of alpha-amanitin (0.7 micrograms/ml) indicating that the T-DNA, although from bacterial origin, is transcribed by the host RNA polymerase II.

MeSH Terms

Amanitins/pharmacology Base Sequence Cell Transformation, Neoplastic DNA/metabolism DNA-Directed RNA Polymerases/metabolism Dactinomycin/pharmacology Nucleic Acid Hybridization Osmolar Concentration Plant Tumors/analysis Plasmids RNA/biosynthesis Transcription, Genetic/drug effects

Chemicals

Amanitins Dactinomycin RNA DNA DNA-Directed RNA Polymerases

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Willmitzer L

Schmalenbach W

Schell J

References (23)

23 references, click to expand

The functional organization of the nopaline A. tumefaciens plasmid pTiC58.
Plasmid. 1980 Mar;3(2):212-30 PMID: 6100894
RNA synthesis in isolated nuclei: in vitro initiation of adenovirus 2 major late mRNA precursor.
Proc Natl Acad Sci U S A. 1979 Jan;76(1):160-4 PMID: 284328
Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.
Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66 PMID: 4894690
Role of DNA-dependent RNA polymerases II and III in transcription of the adenovirus genome late in productive infection.
Proc Natl Acad Sci U S A. 1974 Sep;71(9):3426-39 PMID: 4530313
Inhibition by -amanitin of simian virus 40-specific ribonucleic acid synthesis in nuclei of infected monkey cells.
J Virol. 1972 Nov;10(5):1086-9 PMID: 4344254
Integration and organization of Ti plasmid sequences in crown gall tumors.
Cell. 1980 Mar;19(3):729-39 PMID: 7363328
Transcription of Ti plasmid-derived sequences in three octopine-type crown gall tumor lines.
Proc Natl Acad Sci U S A. 1979 Jun;76(6):2828-32 PMID: 379864
The in vitro synthesis of avian myeloblastosis viral RNA sequences.
Proc Natl Acad Sci U S A. 1974 Aug;71(8):3045-9 PMID: 4370472
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
Nuclear and polysomal transcripts of T-DNA in octopine crown gall suspension and callus cultures.
Mol Gen Genet. 1981;182(2):255-62 PMID: 6945473
In vitro synthesis of Rous sarcoma virus-specific RNA is catalyzed by a DNA-dependent RNA polymerase.
Proc Natl Acad Sci U S A. 1974 Jul;71(7):2782-6 PMID: 4368801
Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis.
Cell. 1977 Jun;11(2):263-71 PMID: 890735
Plasmid required for virulence of Agrobacterium tumefaciens.
J Bacteriol. 1975 Jul;123(1):255-64 PMID: 1141196
A RNA-dependent RNA polymerase activity: implications for chromatin transcription experiments.
Nucleic Acids Res. 1977 Nov;4(11):3943-58 PMID: 339205
Large plasmid in Agrobacterium tumefaciens essential for crown gall-inducing ability.
Nature. 1974 Nov 8;252(5479):169-70 PMID: 4419109
Analysis of in vitro transcription of duck reticulocyte chromatin using mercury-substituted ribonucleoside triphosphates.
Biochemistry. 1977 Nov 15;16(23):5135-45 PMID: 334258
Transcription of viral genes by RNA polymerase II in nuclei isolated from adenovirus 2 transformed cells.
Biochemistry. 1978 May 30;17(11):2198-205 PMID: 667018
Supercoiled circular DNA in crown-gall inducing Agrobacterium strains.
J Mol Biol. 1974 Jun 15;86(1):109-27 PMID: 4854526
Interactions and DNA transfer between Agrobacterium tumefaciens, the Ti-plasmid and the plant host.
Proc R Soc Lond B Biol Sci. 1979 Apr 11;204(1155):251-66 PMID: 36626
Tumor DNA structure in plant cells transformed by A. tumefaciens.
Science. 1980 Sep 19;209(4463):1385-91 PMID: 6251546
T-DNA from Agrobacterium Ti plasmid is in the nuclear DNA fraction of crown gall tumor cells.
Proc Natl Acad Sci U S A. 1980 Jul;77(7):4060-4 PMID: 16592850
Eukaryotic RNA polymerase: comparative subunit structures, immunological properties, and alpha-amanitin sensitivities of the class II enzymes from higher plants.
Biochemistry. 1978 Apr 4;17(7):1322-7 PMID: 656391
Octopine and nopaline synthesis and breakdown genetically controlled by a plasmid of Agrobacterium tumefaciens.
Mol Gen Genet. 1976 May 7;145(2):177-81 PMID: 934053

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/9.19.4801

Published

1981-10-10

Pages

4801-12

Language

English

Region

England

NLM ID

0411011

PMCID

PMC327480

Subset

External Links

PubMed source DOI: 10.1093/nar/9.19.4801 PMC Full Text View journal details

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Transcription of T-DNA in octopine and nopaline crown gall tumours is inhibited by low concentrations of alpha-amanitin.

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