Abstract
A crude membrane-bound RNA polymerase, obtained by differential centrifugation of extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L), was purified by sucrose density gradient centrifugation. Removal of the endogenous RNA template with micrococcal nuclease rendered the polymerase template dependent and template specific. The polymerase was primer independent and able to initiate RNA synthesis on templates containing the 3'-terminal sequences of the TMV-L positive or negative strands. TMV-vulgare RNA was a less efficient template, while RNAs of cucumber mosaic cucumovirus and red clover necrotic mosaic dianthovirus, or 5'-terminal sequences of TMV-L positive or negative strands, did not act as templates for the polymerase. A main product of the reaction with TMV-L genomic RNA as a template, carried out in the presence of [alpha-32P]UTP, was genomic-length single-stranded RNA. This was shown to be the positive strand and uniformly labelled along its length, demonstrating complete replication of TMV-L RNA. Genomic-length double-stranded RNA, labelled in both strands, and small amounts of RNAs corresponding to the single- and double-stranded forms of the coat protein subgenomic mRNA were also formed. Antibodies to N-terminal and C-terminal portions of the 126-kDa protein detected the 126-kDa protein and the 183-kDa readthrough protein in purified RNA polymerase preparations, whereas antibodies to the readthrough portion of the 183-kDa protein detected only the 183-kDa protein. All three antibodies inhibited the template-dependent RNA polymerase, but none of them had any effect on the template-bound enzyme.
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Claims 11
Claim · model-written summary
Removal of endogenous RNA with micrococcal nuclease made the tomato-leaf membrane polymerase template dependent and template specific.
- Taxon
- tomato
Solanum lycopersicume0 - Treatment
- micrococcal nucleasee1
- Interactor
- tobacco mosaic tobamoviruse0pathogen
- Outcome
- template dependent and template specifice1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e1“Removal of the endogenous RNA template with micrococcal nuclease rendered the polymerase template dependent and template specific”
Claim · model-written summary
The membrane polymerase was primer independent and initiated RNA synthesis on TMV-L 3'-terminal positive or negative strand templates.
- Taxon
- tomatoSolanum lycopersicume0
- Interactor
- tobacco mosaic tobamoviruse0pathogen
- Outcome
- primer independente2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e2“The polymerase was primer independent and able to initiate RNA synthesis on templates containing the 3'-terminal sequences of the TMV-L positive or negative strands”
Claim · model-written summary
TMV-vulgare RNA was a less efficient template for the polymerase.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- TMV-vulgare RNAe3
- Outcome
- efficient templatee3
- Direction
- less efficiente3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e3“TMV-vulgare RNA was a less efficient template”
Claim · model-written summary
RNAs of cucumber mosaic cucumovirus and red clover necrotic mosaic dianthovirus did not act as templates for the polymerase.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- RNAs of cucumber mosaic cucumovirus and red clover necrotic mosaic dianthoviruse4
- Outcome
- act as templatese4
- Direction
- did not act as templatese4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e4“RNAs of cucumber mosaic cucumovirus and red clover necrotic mosaic dianthovirus, or 5'-terminal sequences of TMV-L positive or negative strands, did not act as templates for the polymerase”
Claim · model-written summary
A main product of the reaction with TMV-L genomic RNA as template was genomic-length single-stranded RNA.
- Taxon
- tomatoSolanum lycopersicume0
- Outcome
- genomic-length single-stranded RNAe5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e5“A main product of the reaction with TMV-L genomic RNA as a template, carried out in the presence of [alpha-32P]UTP, was genomic-length single-stranded RNA”
Claim · model-written summary
The reaction demonstrated complete replication of TMV-L RNA, with a positive-strand product uniformly labelled along its length.
- Taxon
- tomatoSolanum lycopersicume0
- Interactor
- tobacco mosaic tobamoviruse0pathogen
- Outcome
- complete replication of TMV-L RNAe6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e6“This was shown to be the positive strand and uniformly labelled along its length, demonstrating complete replication of TMV-L RNA”
Claim · model-written summary
Genomic-length double-stranded RNA labelled in both strands was formed, with small amounts of subgenomic RNA forms.
- Taxon
- tomatoSolanum lycopersicume0
- Outcome
- Genomic-length double-stranded RNAe7
- Qualifier
- small amountse7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e7“Genomic-length double-stranded RNA, labelled in both strands, and small amounts of RNAs corresponding to the single- and double-stranded forms of the coat protein subgenomic mRNA were also formed”
Claim · model-written summary
Antibodies to the 126-kDa protein detected the 126-kDa and 183-kDa readthrough proteins in purified polymerase preparations.
- Taxon
- tomatoSolanum lycopersicume0
- Gene
- 126-kDa proteine7
- Outcome
- detected the 126-kDa protein and the 183-kDa readthrough proteine7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e7“Antibodies to N-terminal and C-terminal portions of the 126-kDa protein detected the 126-kDa protein and the 183-kDa readthrough protein in purified RNA polymerase preparations”
Claim · model-written summary
Antibodies to the readthrough portion of the 183-kDa protein detected only the 183-kDa protein.
- Taxon
- tomatoSolanum lycopersicume0
- Gene
- 183-kDa proteine7
- Outcome
- detected only the 183-kDa proteine7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e7“antibodies to the readthrough portion of the 183-kDa protein detected only the 183-kDa protein”
Claim · model-written summary
All three antibodies inhibited the template-dependent RNA polymerase.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- antibodiese7
- Outcome
- template-dependent RNA polymerasee7
- Direction
- inhibitede7
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e7“All three antibodies inhibited the template-dependent RNA polymerase, but none of them had any effect on the template-bound enzyme”
Claim · model-written summary
None of the three antibodies had any effect on the template-bound polymerase.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- antibodiese7
- Outcome
- template-bound enzymee7
- Direction
- had any effecte7
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L)”
e7“All three antibodies inhibited the template-dependent RNA polymerase, but none of them had any effect on the template-bound enzyme”