Structure of the super-elongation complex subunit AFF4 C-terminal homology domain reveals requirements for AFF homo- and heterodimerization.

Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE

Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology

Transcriptional Elongation Factors/*chemistry; Amino Acid Sequence ; Crystallography, X-Ray ; DNA/chemistry ; DNA/metabolism ; Dimerization ; Humans ; Phosphorylation ; Positive Transcriptional Elongation Factor B/metabolism ; Protein Folding ; Protein Structure, Tertiary ; Protein Subunits/chemistry ; Protein Subunits/metabolism ; RNA/chemistry ; RNA/metabolism ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Sequence Alignment ; Substrate Specificity ; Transcriptional Elongation Factors/genetics ; Transcriptional Elongation Factors/metabolism

AF4/FMR2 family member 4 (AFF4) is the scaffold protein of the multisubunit super-elongation complex, which plays key roles in the release of RNA polymerase II from promoter-proximal pausing and in the transactivation of HIV-1 transcription. AFF4 consists of an intrinsically disordered N-terminal region that interacts with other super-elongation complex subunits and a C-terminal homology domain (CHD) that is conserved among AF4/FMR2 family proteins, including AFF1, AFF2, AFF3, and AFF4. Here, we solved the X-ray crystal structure of the CHD in human AFF4 (AFF4-CHD) to 2.2 Å resolution and characterized its biochemical properties. The structure disclosed that AFF4-CHD folds into a novel domain that consists of eight helices and is distantly related to tetratrico peptide repeat motifs. Our analyses further revealed that AFF4-CHD mediates the formation of an AFF4 homodimer or an AFF1-AFF4 heterodimer. Results from fluorescence anisotropy experiments suggested that AFF4-CHD interacts with both RNA and DNA in vitro Furthermore, we identified a surface loop region in AFF4-CHD as a substrate for the P-TEFb kinase cyclin-dependent kinase 9, which triggers release of polymerase II from promoter-proximal pausing sites. In conclusion, the AFF-CHD structure and biochemical analyses reported here reveal the molecular basis for the homo- and heterodimerization of AFF proteins and implicate the AFF4-CHD in nucleic acid interactions. The high conservation of the CHD among several other proteins suggests that our results are also relevant for understanding other CHD-containing proteins and their dimerization behavior.
(© 2019 Chen and Cramer.)

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Keywords: AF4/FMR2 family member 4 (AFF4); C-terminal homology domain (CHD); RNA polymerase II; X-ray crystallography; dimerization; gene regulation; phosphorylation; super elongation complex; transcription elongation factor

PDB 6GKG-F

0 (AFF4 protein, human)
0 (Protein Subunits)
0 (Recombinant Proteins)
0 (Transcriptional Elongation Factors)
63231-63-0 (RNA)
9007-49-2 (DNA)
EC 2.7.11.- (Positive Transcriptional Elongation Factor B)

Date Created: 20190601 Date Completed: 20200310 Latest Revision: 20240720

20240720

PMC6615702

10.1074/jbc.RA119.008577

31147444