Abstract
Some DNA-dependent RNA polymerases (DdRPs) possess RNA-dependent RNA polymerase activity, as was first discovered in the replication of Potato spindle tuber viroid (PSTVd) RNA genome in tomato (Solanum lycopersicum). Recent studies revealed that this activity in bacteria and mammals is important for transcriptional and posttranscriptional regulatory mechanisms. Here, we used PSTVd as a model to uncover auxiliary factors essential for RNA-templated transcription by DdRP PSTVd replication in the nucleoplasm generates (-)-PSTVd intermediates and (+)-PSTVd copies. We found that the Nicotiana benthamiana canonical 9-zinc finger (ZF) Transcription Factor IIIA (TFIIIA-9ZF) as well as its variant TFIIIA-7ZF interacted with (+)-PSTVd, but only TFIIIA-7ZF interacted with (-)-PSTVd. Suppression of TFIIIA-7ZF reduced PSTVd replication, and overexpression of TFIIIA-7ZF enhanced PSTVd replication in planta. Consistent with the locale of PSTVd replication, TFIIIA-7ZF was found in the nucleoplasm and nucleolus, in contrast to the strictly nucleolar localization of TFIIIA-9ZF. Footprinting assays revealed that only TFIIIA-7ZF bound to a region of PSTVd critical for initiating transcription. Furthermore, TFIIIA-7ZF strongly enhanced the in vitro transcription of circular (+)-PSTVd by partially purified Pol II. Together, our results identify TFIIIA-7ZF as a dedicated cellular transcription factor that acts in DdRP-catalyzed RNA-templated transcription, highlighting both the extraordinary evolutionary adaptation of viroids and the potential of DdRPs for a broader role in cellular processes.
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Claims 9
Claim · model-written summary
TFIIIA-9ZF interacted with positive-strand PSTVd.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Interactor
- PSTVde0pathogen
- Gene
- TFIIIA-9ZFe0
- Outcome
- interacted with (+)-PSTVde0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the Nicotiana benthamiana canonical 9-zinc finger (ZF) Transcription Factor IIIA (TFIIIA-9ZF) as well as its variant TFIIIA-7ZF interacted with (+)-PSTVd”
Claim · model-written summary
TFIIIA-7ZF interacted with positive-strand PSTVd.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Interactor
- PSTVde0pathogen
- Gene
- TFIIIA-7ZFe0
- Outcome
- interacted with (+)-PSTVde0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the Nicotiana benthamiana canonical 9-zinc finger (ZF) Transcription Factor IIIA (TFIIIA-9ZF) as well as its variant TFIIIA-7ZF interacted with (+)-PSTVd”
Claim · model-written summary
Only TFIIIA-7ZF interacted with negative-strand PSTVd.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Interactor
- PSTVde0pathogen
- Gene
- TFIIIA-7ZFe0
- Outcome
- interacted with (-)-PSTVde0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“but only TFIIIA-7ZF interacted with (-)-PSTVd”
e1“Nicotiana benthamiana”
Claim · model-written summary
Suppressing TFIIIA-7ZF reduced PSTVd replication.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- Suppression of TFIIIA-7ZFe1
- Interactor
- PSTVde1pathogen
- Gene
- TFIIIA-7ZFe1
- Outcome
- PSTVd replicatione1
- Direction
- reducede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“Suppression of TFIIIA-7ZF reduced PSTVd replication”
Claim · model-written summary
Overexpressing TFIIIA-7ZF enhanced PSTVd replication in planta.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- overexpression of TFIIIA-7ZFe1
- Interactor
- PSTVde1pathogen
- Gene
- TFIIIA-7ZFe1
- Outcome
- PSTVd replicatione1
- Direction
- enhancede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“overexpression of TFIIIA-7ZF enhanced PSTVd replication in planta”
Claim · model-written summary
TFIIIA-7ZF localized to nucleoplasm and nucleolus.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Gene
- TFIIIA-7ZFe1
- Outcome
- nucleoplasm and nucleoluse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“TFIIIA-7ZF was found in the nucleoplasm and nucleolus”
Claim · model-written summary
TFIIIA-9ZF was strictly nucleolar.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Gene
- TFIIIA-9ZFe1
- Outcome
- strictly nucleolar localizatione1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“strictly nucleolar localization of TFIIIA-9ZF”
Claim · model-written summary
Only TFIIIA-7ZF bound a PSTVd region critical for initiating transcription.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Interactor
- PSTVde1pathogen
- Gene
- TFIIIA-7ZFe1
- Outcome
- bound to a region of PSTVd critical for initiating transcriptione1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“only TFIIIA-7ZF bound to a region of PSTVd critical for initiating transcription”
Claim · model-written summary
TFIIIA-7ZF strongly enhanced in vitro transcription of circular positive-strand PSTVd.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- TFIIIA-7ZFe1
- Interactor
- PSTVde1pathogen
- Gene
- TFIIIA-7ZFe1
- Outcome
- in vitro transcription of circular (+)-PSTVde1
- Direction
- enhancede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana”
e1“TFIIIA-7ZF strongly enhanced the in vitro transcription of circular (+)-PSTVd by partially purified Pol II”
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