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Chemical perturbation of an intrinsically disordered region of TFIID distinguishes two modes of transcription initiation.

Zhang Z, Boskovic Z, Hussain MM, Hu W, Inouye C, Kim HJ, Abole AK, Doud MK, Lewis TA, Koehler AN, Schreiber SL, Tjian R - Elife (2015)

Bottom Line: They are abundant in eukaryotic proteomes and are often associated with human diseases, but their biological functions have been elusive to study.Binding arrests an isomerization of promoter-bound TFIID that is required for the engagement of Pol II during the first (de novo) round of transcription initiation.This work also suggests a new avenue for targeting the elusive IDRs by harnessing certain features of metal-based complexes for mechanistic studies, and for the development of novel pharmaceutical interventions.

View Article: PubMed Central - PubMed

Affiliation: Transcription Imaging Consortium, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States.

ABSTRACT
Intrinsically disordered proteins/regions (IDPs/IDRs) are proteins or peptide segments that fail to form stable 3-dimensional structures in the absence of partner proteins. They are abundant in eukaryotic proteomes and are often associated with human diseases, but their biological functions have been elusive to study. In this study, we report the identification of a tin(IV) oxochloride-derived cluster that binds an evolutionarily conserved IDR within the metazoan TFIID transcription complex. Binding arrests an isomerization of promoter-bound TFIID that is required for the engagement of Pol II during the first (de novo) round of transcription initiation. However, the specific chemical probe does not affect reinitiation, which requires the re-entry of Pol II, thus, mechanistically distinguishing these two modes of transcription initiation. This work also suggests a new avenue for targeting the elusive IDRs by harnessing certain features of metal-based complexes for mechanistic studies, and for the development of novel pharmaceutical interventions.

No MeSH data available.


Related in: MedlinePlus

The intrinsically disordered nature of metazoan TAF2 C-terminus and the stringent conservation of and around the low-complexity sequences.Full-length TAF2 proteins from Drosophila (A) or human (B) were analyzed for protein-disordered region (http://bioinf.cs.ucl.ac.uk/psipred/?disopred=1). The predicted α-helix and β-sheet structures are highlighted in purple and yellow, respectively, and the C-terminus-disordered regions are underscored. (C) Alignment of the very C-terminus of TAF2 proteins from multiple vertebrates. The low-complexity sequences and other well-conserved residues were highlighted in distinct colors.DOI:http://dx.doi.org/10.7554/eLife.07777.011
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fig4s1: The intrinsically disordered nature of metazoan TAF2 C-terminus and the stringent conservation of and around the low-complexity sequences.Full-length TAF2 proteins from Drosophila (A) or human (B) were analyzed for protein-disordered region (http://bioinf.cs.ucl.ac.uk/psipred/?disopred=1). The predicted α-helix and β-sheet structures are highlighted in purple and yellow, respectively, and the C-terminus-disordered regions are underscored. (C) Alignment of the very C-terminus of TAF2 proteins from multiple vertebrates. The low-complexity sequences and other well-conserved residues were highlighted in distinct colors.DOI:http://dx.doi.org/10.7554/eLife.07777.011

Mentions: Identification of tin(IV) oxochloride led us to consider presumptive targets that should be histidine-rich domains of phylogenetically conserved TAF subunits. Testing this hypothesis, we found that a GST fusion of the histidine-rich Drosophila TAF2 (dTAF2) C-terminal fragment (residues [1125–1221]) bound selectively to the original tin-oxochloride containing sample in the arrayed library (Figure 4A). The targeted polypeptide fragment is part of a conserved IDR with adjacent low-complexity poly (K), poly (KH), and poly (KD/E) motifs found in both Drosophila and human TAF2 (Verrijzer et al., 1994; Kaufmann et al., 1998) (Figure 4B and Figure 4—figure supplement 1). The tin(IV) oxochloride cluster, with its hydrophilic, periodic surface features, presents a likely complementary ligand for these polar, repetitive, and histidine-rich IDRs.10.7554/eLife.07777.010Figure 4.The inhibitor targets an IDR of TAF2 through histidines.


Chemical perturbation of an intrinsically disordered region of TFIID distinguishes two modes of transcription initiation.

Zhang Z, Boskovic Z, Hussain MM, Hu W, Inouye C, Kim HJ, Abole AK, Doud MK, Lewis TA, Koehler AN, Schreiber SL, Tjian R - Elife (2015)

The intrinsically disordered nature of metazoan TAF2 C-terminus and the stringent conservation of and around the low-complexity sequences.Full-length TAF2 proteins from Drosophila (A) or human (B) were analyzed for protein-disordered region (http://bioinf.cs.ucl.ac.uk/psipred/?disopred=1). The predicted α-helix and β-sheet structures are highlighted in purple and yellow, respectively, and the C-terminus-disordered regions are underscored. (C) Alignment of the very C-terminus of TAF2 proteins from multiple vertebrates. The low-complexity sequences and other well-conserved residues were highlighted in distinct colors.DOI:http://dx.doi.org/10.7554/eLife.07777.011
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582147&req=5

fig4s1: The intrinsically disordered nature of metazoan TAF2 C-terminus and the stringent conservation of and around the low-complexity sequences.Full-length TAF2 proteins from Drosophila (A) or human (B) were analyzed for protein-disordered region (http://bioinf.cs.ucl.ac.uk/psipred/?disopred=1). The predicted α-helix and β-sheet structures are highlighted in purple and yellow, respectively, and the C-terminus-disordered regions are underscored. (C) Alignment of the very C-terminus of TAF2 proteins from multiple vertebrates. The low-complexity sequences and other well-conserved residues were highlighted in distinct colors.DOI:http://dx.doi.org/10.7554/eLife.07777.011
Mentions: Identification of tin(IV) oxochloride led us to consider presumptive targets that should be histidine-rich domains of phylogenetically conserved TAF subunits. Testing this hypothesis, we found that a GST fusion of the histidine-rich Drosophila TAF2 (dTAF2) C-terminal fragment (residues [1125–1221]) bound selectively to the original tin-oxochloride containing sample in the arrayed library (Figure 4A). The targeted polypeptide fragment is part of a conserved IDR with adjacent low-complexity poly (K), poly (KH), and poly (KD/E) motifs found in both Drosophila and human TAF2 (Verrijzer et al., 1994; Kaufmann et al., 1998) (Figure 4B and Figure 4—figure supplement 1). The tin(IV) oxochloride cluster, with its hydrophilic, periodic surface features, presents a likely complementary ligand for these polar, repetitive, and histidine-rich IDRs.10.7554/eLife.07777.010Figure 4.The inhibitor targets an IDR of TAF2 through histidines.

Bottom Line: They are abundant in eukaryotic proteomes and are often associated with human diseases, but their biological functions have been elusive to study.Binding arrests an isomerization of promoter-bound TFIID that is required for the engagement of Pol II during the first (de novo) round of transcription initiation.This work also suggests a new avenue for targeting the elusive IDRs by harnessing certain features of metal-based complexes for mechanistic studies, and for the development of novel pharmaceutical interventions.

View Article: PubMed Central - PubMed

Affiliation: Transcription Imaging Consortium, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States.

ABSTRACT
Intrinsically disordered proteins/regions (IDPs/IDRs) are proteins or peptide segments that fail to form stable 3-dimensional structures in the absence of partner proteins. They are abundant in eukaryotic proteomes and are often associated with human diseases, but their biological functions have been elusive to study. In this study, we report the identification of a tin(IV) oxochloride-derived cluster that binds an evolutionarily conserved IDR within the metazoan TFIID transcription complex. Binding arrests an isomerization of promoter-bound TFIID that is required for the engagement of Pol II during the first (de novo) round of transcription initiation. However, the specific chemical probe does not affect reinitiation, which requires the re-entry of Pol II, thus, mechanistically distinguishing these two modes of transcription initiation. This work also suggests a new avenue for targeting the elusive IDRs by harnessing certain features of metal-based complexes for mechanistic studies, and for the development of novel pharmaceutical interventions.

No MeSH data available.


Related in: MedlinePlus