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Archaeal TFEα/β is a hybrid of TFIIE and the RNA polymerase III subcomplex hRPC62/39.

Blombach F, Salvadori E, Fouqueau T, Yan J, Reimann J, Sheppard C, Smollett KL, Albers SV, Kay CW, Thalassinos K, Werner F - Elife (2015)

Bottom Line: The eukaryotic transcription factor TFIIE consists of α and β subunits.Here we have identified and characterised the function of the TFIIEβ homologue in archaea that on the primary sequence level is related to the RNAPIII subunit hRPC39.These activities are strictly dependent on the β subunit and the promoter sequence.

View Article: PubMed Central - PubMed

Affiliation: Institute for Structural and Molecular Biology, Division of Biosciences, University College London, London, United Kingdom.

ABSTRACT
Transcription initiation of archaeal RNA polymerase (RNAP) and eukaryotic RNAPII is assisted by conserved basal transcription factors. The eukaryotic transcription factor TFIIE consists of α and β subunits. Here we have identified and characterised the function of the TFIIEβ homologue in archaea that on the primary sequence level is related to the RNAPIII subunit hRPC39. Both archaeal TFEβ and hRPC39 harbour a cubane 4Fe-4S cluster, which is crucial for heterodimerization of TFEα/β and its engagement with the RNAP clamp. TFEα/β stabilises the preinitiation complex, enhances DNA melting, and stimulates abortive and productive transcription. These activities are strictly dependent on the β subunit and the promoter sequence. Our results suggest that archaeal TFEα/β is likely to represent the evolutionary ancestor of TFIIE-like factors in extant eukaryotes.

No MeSH data available.


Related in: MedlinePlus

TFEα/β is part of the PIC.Supershift EMSA with PICs formed in presence or absence of TFEα/β (0.1 µM) being exposed to antisera directed against TFEα, TFEβ and TBP. Antibody-PIC complexes migrate slower in the gel compared to PIC alone causing a supershift.DOI:http://dx.doi.org/10.7554/eLife.08378.017
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fig6s1: TFEα/β is part of the PIC.Supershift EMSA with PICs formed in presence or absence of TFEα/β (0.1 µM) being exposed to antisera directed against TFEα, TFEβ and TBP. Antibody-PIC complexes migrate slower in the gel compared to PIC alone causing a supershift.DOI:http://dx.doi.org/10.7554/eLife.08378.017

Mentions: Deletion of the TFEα ZR or the TFEβ WH domains reduced the stimulation. The former mutation destabilizes the binding to the RNAP, while the latter variant is not impaired in RNAP binding (Figure 5), which suggests that the TFEβ WH domain plays a role for the stabilisation of the PIC. EMSA supershift experiments validated the incorporation of TFEβ into the PIC (Figure 6—figure supplement 1).


Archaeal TFEα/β is a hybrid of TFIIE and the RNA polymerase III subcomplex hRPC62/39.

Blombach F, Salvadori E, Fouqueau T, Yan J, Reimann J, Sheppard C, Smollett KL, Albers SV, Kay CW, Thalassinos K, Werner F - Elife (2015)

TFEα/β is part of the PIC.Supershift EMSA with PICs formed in presence or absence of TFEα/β (0.1 µM) being exposed to antisera directed against TFEα, TFEβ and TBP. Antibody-PIC complexes migrate slower in the gel compared to PIC alone causing a supershift.DOI:http://dx.doi.org/10.7554/eLife.08378.017
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495717&req=5

fig6s1: TFEα/β is part of the PIC.Supershift EMSA with PICs formed in presence or absence of TFEα/β (0.1 µM) being exposed to antisera directed against TFEα, TFEβ and TBP. Antibody-PIC complexes migrate slower in the gel compared to PIC alone causing a supershift.DOI:http://dx.doi.org/10.7554/eLife.08378.017
Mentions: Deletion of the TFEα ZR or the TFEβ WH domains reduced the stimulation. The former mutation destabilizes the binding to the RNAP, while the latter variant is not impaired in RNAP binding (Figure 5), which suggests that the TFEβ WH domain plays a role for the stabilisation of the PIC. EMSA supershift experiments validated the incorporation of TFEβ into the PIC (Figure 6—figure supplement 1).

Bottom Line: The eukaryotic transcription factor TFIIE consists of α and β subunits.Here we have identified and characterised the function of the TFIIEβ homologue in archaea that on the primary sequence level is related to the RNAPIII subunit hRPC39.These activities are strictly dependent on the β subunit and the promoter sequence.

View Article: PubMed Central - PubMed

Affiliation: Institute for Structural and Molecular Biology, Division of Biosciences, University College London, London, United Kingdom.

ABSTRACT
Transcription initiation of archaeal RNA polymerase (RNAP) and eukaryotic RNAPII is assisted by conserved basal transcription factors. The eukaryotic transcription factor TFIIE consists of α and β subunits. Here we have identified and characterised the function of the TFIIEβ homologue in archaea that on the primary sequence level is related to the RNAPIII subunit hRPC39. Both archaeal TFEβ and hRPC39 harbour a cubane 4Fe-4S cluster, which is crucial for heterodimerization of TFEα/β and its engagement with the RNAP clamp. TFEα/β stabilises the preinitiation complex, enhances DNA melting, and stimulates abortive and productive transcription. These activities are strictly dependent on the β subunit and the promoter sequence. Our results suggest that archaeal TFEα/β is likely to represent the evolutionary ancestor of TFIIE-like factors in extant eukaryotes.

No MeSH data available.


Related in: MedlinePlus