Untitled

Kowalski, Kasper; Czolij, R.; King, Glenn F.; Crossley, Merlin; Mackay, Joel P.

doi:10.1023/a:1008309602929

Cited by 50 publications

(23 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As for both of the previously reported zinc fingers of GATA-1 (15,17), the GATA-like finger of the ADD domain comprises an irregular hairpin loop carrying the first two zinc ligands, followed by a short antiparallel ␤-sheet (residues 186-189, s1, paired with residues 194-197, s2) and a short ␣-helix (residues 198-206, h1), the latter starting between the second pair of zinc ligands. The correspondence between the GATA-like finger of the ADD domain and the fingers from GATA-1 itself is very close (Fig.…”

Section: Nmr Structurementioning

confidence: 84%

Structural consequences of disease-causing mutations in the ATRX-DNMT3-DNMT3L (ADD) domain of the chromatin-associated protein ATRX

Argentaro

Yang

Chapman

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

140

120

View full text Add to dashboard Cite

The chromatin-associated protein ATRX was originally identified because mutations in the ATRX gene cause a severe form of syndromal X-linked mental retardation associated with ␣-thalassemia. Half of all of the disease-associated missense mutations cluster in a cysteine-rich region in the N terminus of ATRX. This region was named the ATRX-DNMT3-DNMT3L (ADD) domain, based on sequence homology with a family of DNA methyltransferases. Here, we report the solution structure of the ADD domain of ATRX, which consists of an N-terminal GATA-like zinc finger, a plant homeodomain finger, and a long C-terminal ␣-helix that pack together to form a single globular domain. Interestingly, the ␣-helix of the GATA-like finger is exposed and highly basic, suggesting a DNA-binding function for ATRX. The disease-causing mutations fall into two groups: the majority affect buried residues and hence affect the structural integrity of the ADD domain; another group affects a cluster of surface residues, and these are likely to perturb a potential protein interaction site. The effects of individual point mutations on the folding state and stability of the ADD domain correlate well with the levels of mutant ATRX protein in patients, providing insights into the molecular pathophysiology of ATR-X syndrome.ATR-X syndrome ͉ NMR structure ͉ zinc finger A TRX was identified when the gene encoding this protein was shown to be mutated in a form of X-linked mental retardation (ATR-X syndrome) in young males (1, 2). Furthermore, null mutations in mice are lethal at the embryonic stage of development (3). Because ATRX mutations reduce ␣-globin synthesis, causing ␣-thalassemia, it seems likely that ATRX normally plays a role in the regulation of globin gene expression (2, 4). The complexity of the disease also suggests that ATRX could be involved in the regulation of other as yet unidentified genes. ATRX is a large (2,492 residue; Ϸ280 kDa) nuclear protein predominantly localized to heterochromatin and nuclear PML bodies (5, 6). It contains two highly conserved domains, and missense mutations that give rise to ATR-X syndrome fall within these. At the C terminus is a helicase/ATPase domain, which characterizes ATRX as a member of the SNF2 (SWI/SNF) family of chromatin-associated proteins. Experimental evidence shows that ATRX acts as a DNA-dependent ATPase and as a DNA translocase, and it confers modest chromatin-remodeling activity in vitro (6). Thus, it seems likely that ATRX exerts its function by targeting chromatin.Of the missense mutations identified in the ATRX gene, 50% are located in the N terminus of the ATRX protein, which represents just 4% of the coding sequence (Fig. 1a) (7). This region is highly cysteine-rich and contains two different types of zinc-finger motif. It was first noticed that a region of the sequence shares homology with the plant homeodomain (PHD)-type zinc fingers (8). Mutations in other PHD-containing proteins (WSTF and AIRE) are also associated with human disease (9, 10). PHD fingers are found in nuclear proteins, and a...

show abstract

Section: Nmr Structurementioning

confidence: 84%

Structural consequences of disease-causing mutations in the ATRX-DNMT3-DNMT3L (ADD) domain of the chromatin-associated protein ATRX

Argentaro

Yang

Chapman

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

140

120

View full text Add to dashboard Cite

show abstract

“…Valine-205 resides in the N-terminal finger on the surface opposite to that involved in DNA binding and contacts FOG-1 but not DNA (29). Replacement of V205 with methionine or glycine diminishes FOG-1 binding without affecting the interaction with DNA in vitro (13,14,30).…”

Section: Fog-1 Requirement For Gata-1-induced Histone Acetylation At mentioning

confidence: 99%

Context-dependent regulation of GATA-1 by friend of GATA-1

Letting

Chen²,

Rakowski³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

126

134

View full text Add to dashboard Cite

The transcription factor GATA-1 and its cofactor, friend of GATA-1 (FOG-1), are essential for normal erythroid development. FOG-1 physically interacts with GATA-1 to augment or inhibit its activity. The mechanisms by which FOG-1 regulates GATA-1 function are unknown. By using an assay that is based on the phenotypic rescue of a GATA-1-null erythroid cell line, we found that a conditional form of GATA-1 (GATA-1-ER) strongly induced histone acetylation at the ␤-major globin promoter in vivo, consistent with previous results. In contrast, GATA-1 bearing a point mutation that impairs FOG-1 binding [GATA-1(V205M)-ER] failed to induce high levels of histone acetylation at this site. However, at DNase I-hypersensitive site (HS)3 of the ␤-globin locus control region, GATA-1-induced histone acetylation was FOG-1-independent. Because the V205M mutation does not disrupt GATA-1 binding to DNA templates in vitro, we were surprised to find that in vivo GATA-1(V205M)-ER fails to bind the ␤-globin promoter. However, at HS3, DNA binding by GATA-1 was FOG-1-independent, thus correlating histone acetylation with GATA-1 occupancy. Examination of additional GATA-1-dependent regulatory elements showed that the interaction with FOG-1 is required for GATA-1 occupancy at select sites, such as HS2, but is dispensable at others, including the FOG-1-independent GATA-1 target gene EKLF. Remarkably, at the GATA-2 gene, which is repressed by GATA-1, interaction with FOG-1 was dispensable for GATA-1 occupancy and was required for transcriptional inhibition and histone deacetylation. These results indicate that FOG-1 employs distinct mechanisms when cooperating with GATA-1 during transcriptional activation and repression.globin ͉ chromatin ͉ histones ͉ acetylation

show abstract

“…Expression and Purification-USH-F1, USH-F9, and murine GATA-1 NF were expressed and purified as previously described (16,23). Pannier NF (Pannier (165-208)) was subcloned, overexpressed, and purified using the same protocol as that used for the GATA-1 NF, except that the template for PCR of the construct was obtained from a D. melanogaster cDNA library.…”

Section: Methodsmentioning

confidence: 99%

“…The amplified oligonucleotide was inserted into the Escherichia coli expression vector pGEX-2T (Amersham Biosciences). The peptide (with residues numbered 1-36, including Gly-1 and Ser-2, which are the product of the thrombin recognition sequence) was overexpressed and purified by glutathione-affinity chromatography and reverse-phase high performance liquid chromatography as previously described (23). The purification yielded ϳ2 mg of Ͼ95% pure peptide per liter of culture.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of the Conserved Interaction between GATA and FOG Family Proteins

Kowalski¹,

Liew²,

Matthews³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

The N-terminal zinc finger (ZnF) from GATA transcription factors mediates interactions with FOG family proteins. In FOG proteins, the interacting domains are also ZnFs; these domains are related to classical CCHH fingers but have an His 3 Cys substitution at the final zinc-ligating position. Here we demonstrate that different CCHC fingers in the FOG family protein U-shaped contact the N-terminal ZnF of GATA-1 in the same fashion although with different affinities. We also show that these interactions are of moderate affinity, which is interesting given the presumed low concentrations of these proteins in the nucleus. Furthermore, we demonstrate that the variant CCHC topology enhances binding affinity, although the His 3 Cys change is not essential for the formation of a stably folded domain. To ascertain the structural basis for the contribution of the CCHC arrangement, we have determined the structure of a CCHH mutant of finger nine from U-shaped. The structure is very similar overall to the wild-type domain, with subtle differences at the C terminus that result in loss of the interaction in vivo. Taken together, these results suggest that the CCHC zinc binding topology is required for the integrity of GATA-FOG interactions and that weak interactions can play important roles in vivo.

show abstract

Untitled

Cited by 50 publications

References 48 publications

Structural consequences of disease-causing mutations in the ATRX-DNMT3-DNMT3L (ADD) domain of the chromatin-associated protein ATRX

Structural consequences of disease-causing mutations in the ATRX-DNMT3-DNMT3L (ADD) domain of the chromatin-associated protein ATRX

Context-dependent regulation of GATA-1 by friend of GATA-1

Characterization of the Conserved Interaction between GATA and FOG Family Proteins

Contact Info

Product

Resources

About