Expression and nuclear localization of the TATA-box-binding protein during baculovirus infection

The TATA-box-binding protein (TBP) plays a key role in initiating eukaryotic transcription and is used by many viruses for viral transcription. We previously reported increased TBP levels during infection with the baculovirus Autographa californica multicapsid nuclear polyhedrovirus (AcMNPV). The TBP antiserum used in that study, however, cross-reacted with a baculoviral protein. Here, we reported that increased amounts of nuclear TBP were detected upon infection of Spodoptera frugiperda and TN-368 cells with a TBP-specific antiserum. TBP levels increased until 72 h post-infection (p.i.), whilst tbp transcripts decreased by 16 h p.i., which suggested a virus-induced influence on the TBP protein levels. To address a potential modification of the TBP degradation pathway during infection, we investigated the possible role of viral ubiquitin. Infection studies with AcMNPV recombinants carrying a mutated viral ubiquitin gene revealed that the TBP increase during infection was not altered. In addition, pulse–chase experiments indicated a high TBP half-life of ~60 h in uninfected cells, suggesting that a virus-induced increase of TBP stability was unlikely. This increase in TBP correlated with a redistribution to nuclear domains resembling sites of viral DNA synthesis. Furthermore, we observed colocalization of TBP with host RNA polymerase (RNAP) II, but only until 8 h p.i., whilst TBP, but not RNAPII, was present in the enlarged replication domains late during infection. Thus, we suggested that AcMNPV adapted a mechanism to accumulate the highly stable cellular TBP at sites of viral DNA replication and transcription.

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The gammaherpesviral TATA-box-binding protein directly interacts with the CTD of host RNA Pol II to direct late gene transcription

10.1101/2020.01.28.922526 ◽

2020 ◽

Author(s):

Angelica F. Castañeda ◽

Allison L. Didychuk ◽

Robert K. Louder ◽

Chloe O. McCollum ◽

Zoe H. Davis ◽

...

Keyword(s):

Rna Polymerase Ii ◽

Gene Transcription ◽

Binding Protein ◽

Tata Box ◽

Late Gene ◽

Rna Pol Ii ◽

Eukaryotic Transcription ◽

Pol Ii ◽

Terminal Domain ◽

Tata Box Binding Protein

ABSTRACTβ- and γ-herpesviruses include the oncogenic human viruses Kaposi’s sarcoma-associated virus (KSHV) and Epstein-Barr virus (EBV), and human cytomegalovirus (HCMV), which is a significant cause of congenital disease. Near the end of their replication cycle, these viruses transcribe their late genes in a manner distinct from host transcription. Late gene transcription requires six virally-encoded proteins, one of which is a functional mimic of host TATA-box-binding protein (TBP) that is also involved in recruitment of RNA polymerase II (Pol II) via unknown mechanisms. Here, we applied biochemical protein interaction studies together with electron microscopy-based imaging of a reconstituted human preinitiation complex to define the mechanism underlying Pol II recruitment. These data revealed that the herpesviral TBP, encoded by ORF24 in KSHV, makes a direct protein-protein contact with the C-terminal domain of host RNA polymerase II (Pol II), which is a unique feature that functionally distinguishes viral from cellular TBP. The interaction is mediated by the N-terminal domain (NTD) of ORF24 through a conserved motif that is shared in its β- and γ-herpesvirus homologs. Thus, these herpesviruses employ an unprecedented strategy in eukaryotic transcription, wherein promoter recognition and polymerase recruitment are facilitated by a single transcriptional activator with functionally distinct domains.SIGNIFICANCE STATEMENTThe β- and γ-herpesviruses mediate their late gene transcription through a set of viral transcriptional activators (vTAs). One of these vTAs, ORF24 in Kaposi’s sarcoma-associated herpesvirus (KSHV), is a mimic of host TATA-box-binding protein (TBP). We demonstrate that the N-terminal domain of ORF24 and its homologs from other β- and γ-herpesviruses directly bind the unstructured C-terminal domain (CTD) of RNA Pol II. This functionally distinguishes the viral TBP mimic from cellular TBP, which does not bind Pol II. Thus, herpesviruses encode a transcription factor that has the dual ability to directly interact with promoter DNA and the polymerase, a property which is unique in eukaryotic transcription and is conceptually akin to prokaryotic transcription factors.

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Faculty Opinions recommendation of The VirD2 pilot protein of Agrobacterium-transferred DNA interacts with the TATA box-binding protein and a nuclear protein kinase in plants.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1014940.195136 ◽

2003 ◽

Author(s):

Steven Spiker

Keyword(s):

Protein Kinase ◽

Nuclear Protein ◽

Binding Protein ◽

Tata Box ◽

Tata Box Binding Protein

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Identification of the cis-acting DNA sequence elements regulating the transcription of the Saccharomyces cerevisiae gene encoding TBP, the TATA box binding protein.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)46933-5 ◽

1994 ◽

Vol 269 (45) ◽

pp. 28335-28346

Author(s):

S C Schroeder ◽

C K Wang ◽

P A Weil

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Sequence ◽

Binding Protein ◽

Tata Box ◽

Gene Encoding ◽

Cis Acting ◽

Sequence Elements ◽

Tata Box Binding Protein

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Ectopic expression of TATA box-binding protein induces shoot proliferation in Arabidopsis

FEBS Letters ◽

10.1016/s0014-5793(01)02101-9 ◽

2001 ◽

Vol 489 (2-3) ◽

pp. 187-191 ◽

Cited By ~ 9

Author(s):

You-Fang Li ◽

Frédéric Dubois ◽

Dao-Xiu Zhou

Keyword(s):

Binding Protein ◽

Ectopic Expression ◽

Shoot Proliferation ◽

Tata Box ◽

Tata Box Binding Protein

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Bidirectional binding of the TATA box binding protein to the TATA box

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.94.25.13475 ◽

1997 ◽

Vol 94 (25) ◽

pp. 13475-13480 ◽

Cited By ~ 51

Author(s):

J. M. Cox ◽

M. M. Hayward ◽

J. F. Sanchez ◽

L. D. Gegnas ◽

S. van der Zee ◽

...

Keyword(s):

Binding Protein ◽

Tata Box ◽

Tata Box Binding Protein

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Topological localization of the human transcription factors IIA, IIB, TATA box-binding protein, and RNA polymerase II-associated protein 30 on a class II promoter.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)32114-2 ◽

1994 ◽

Vol 269 (31) ◽

pp. 19962-19967 ◽

Cited By ~ 2

Author(s):

B. Coulombe ◽

J. Li ◽

J. Greenblatt

Keyword(s):

Transcription Factors ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Binding Protein ◽

Class Ii ◽

Tata Box ◽

Topological Localization ◽

Tata Box Binding Protein

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c-Fos-induced activation of a TATA-box-containing promoter involves direct contact with TATA-box-binding protein

Molecular and Cellular Biology ◽

10.1128/mcb.14.9.6021-6029.1994 ◽

1994 ◽

Vol 14 (9) ◽

pp. 6021-6029

Author(s):

R Metz ◽

A J Bannister ◽

J A Sutherland ◽

C Hagemeier ◽

E C O'Rourke ◽

...

Keyword(s):

Protein Interactions ◽

Transcriptional Activation ◽

Binding Protein ◽

Tata Box ◽

Binding Motif ◽

Protein Protein Interactions ◽

High Concentrations ◽

Tata Box Binding Protein

Transcriptional activation in eukaryotes involves protein-protein interactions between regulatory transcription factors and components of the basal transcription machinery. Here we show that c-Fos, but not a related protein, Fra-1, can bind the TATA-box-binding protein (TBP) both in vitro and in vivo and that c-Fos can also interact with the transcription factor IID complex. High-affinity binding to TBP requires c-Fos activation modules which cooperate to activate transcription. One of these activation modules contains a TBP-binding motif (TBM) which was identified through its homology to TBP-binding viral activators. This motif is required for transcriptional activation, as well as TBP binding. Domain swap experiments indicate that a domain containing the TBM can confer TBP binding on Fra-1 both in vitro and in vivo. In vivo activation experiments indicate that a GAL4-Fos fusion can activate a promoter bearing a GAL4 site linked to a TATA box but that this activity does not occur at high concentrations of GAL4-Fos. This inhibition (squelching) of c-Fos activity is relieved by the presence of excess TBP, indicating that TBP is a direct functional target of c-Fos. Removing the TBM from c-Fos severely abrogates activation of a promoter containing a TATA box but does not affect activation of a promoter driven only by an initiator element. Collectively, these results suggest that c-Fos is able to activate via two distinct mechanisms, only one of which requires contact with TBP. Since TBP binding is not exhibited by Fra-1, TBP-mediated activation may be one characteristic that discriminates the function of Fos-related proteins.

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