Critical YxKxHxxxRP Motif in the C-Terminal Region of GATA3 for Its DNA Binding and Function

Abstract

A zinc finger transcription factor, GATA3, plays an essential role in the development of T cells and the functional differentiation into type 2 Th cells. Two transactivation domains and two zinc finger regions are known to be important for the GATA3 function, whereas the role for other regions remains unclear. In this study we demonstrated that a conserved YxKxHxxxRP motif (aa 345–354) adjacent to the C-terminal zinc finger domain of GATA3 plays a critical in its DNA binding and functions, including transcriptional activity, the ability to induce chromatin remodeling of the Th2 cytokine gene loci, and Th2 cell differentiation. A single point mutation of the key amino acid (Y, K, H, R, and P) in the motif abrogated GATA3 functions. A computer simulation analysis based on the solution structure of the chicken GATA1/DNA complex supported the importance of this motif in GATA3 DNA binding. Thus, we identified a novel conserved YxKxHxxxRP motif adjacent to the C-terminal zinc finger domain of GATA3 that is indispensable for GATA3 DNA binding and functions.

Genome-Wide Analysis Reveals Unique Regulation of Transcription of Th2-Specific Genes by GATA3

“…The pMX-IRES-human nerve growth factor receptor (NGFR) plasmid was generated, as previously described (16). The infected cells were enriched FIGURE 1.…”

“…In peripheral CD4 T cells, the activation of STAT6 induces highlevel GATA3 mRNA expression (12), and we and others have recently reported that STAT6 binds to specific regions of the GATA3 gene to regulate its transcription (13,14). In addition, the protein expression of GATA3 in Th2 cells is regulated by various posttranscriptional mechanisms (15)(16)(17). Changes in histone modifications, such as H3-K9/14 acetylation and H3-K4 methylation at the Th2 cytokine gene loci, occur during Th2 cell differentiation (18)(19)(20), and this is induced primarily by GATA3 in both CD4 and CD8 T cells (21).…”

Identification of a New Pathway for Th1 Cell Development Induced by Cooperative Stimulation with IL-4 and TGF-β

“…The pMX-IRES-hNGFR plasmid was generated, as described previously (26). Retrovirus vectors containing dominant-negative Eomes (Eomes-DN) cDNA (27) (27).…”

The Interplay between Transcription Factors and Epigenetic Modifications in Th2 Cells

“…Enforced expression of GATA3 has also been reported to endogenously upregulate GATA3 expression [36]. In addition, the amount of GATA3 protein in Th2 cells is regulated by various postranscriptional mechanisms [37][38][39]. Furthermore, high-level expression of GATA3 is essential for the production of large amounts of Th2 cytokines in established Th2 cells [11,[40][41][42].…”