The transcriptional co-activator TAZ interacts differentially with transcriptional enhancer factor-1 (TEF-1) family members

Abstract

Members of the highly related TEF-1 (transcriptional enhancer factor-1) family (also known as TEAD, for TEF-1, TEC1, ABAA domain) bind to MCAT (muscle C, A and T sites) and A/T-rich sites in promoters active in cardiac, skeletal and smooth muscle, placenta, and neural crest. TEF-1 activity is regulated by interactions with transcriptional co-factors [p160, TONDU (Vgl-1, Vestigial-like protein-1), Vgl-2 and YAP65 (Yes-associated protein 65 kDa)]. The strong transcriptional co-activator YAP65 interacts with all TEF-1 family members, and, since YAP65 is related to TAZ (transcriptional co-activator with PDZ-binding motif), we wanted to determine if TAZ also interacts with members of the TEF-1 family. In the present study, we show by GST (glutathione S-transferase) pull-down assays, by co-immunoprecipitation and by modified mammalian two-hybrid assays that TEF-1 interacts with TAZ in vitro and in vivo. Electrophoretic mobility-shift assays with purified TEF-1 and GST-TAZ fusion protein showed that TAZ interacts with TEF-1 bound to MCAT DNA. TAZ can interact with endogenous TEF-1 proteins, since exogenous TAZ activated MCAT-dependent reporter promoters. Like YAP65, TAZ interacted with all four TEF-1 family members. GST pull-down assays with increasing amounts of [ 35 S]TEF-1 and [ 35 S]RTEF-1 (related TEF-1) showed that TAZ interacts more efficiently with TEF-1 than with RTEF-1. This differential interaction also extended to the interaction of TEF-1 and RTEF-1 with TAZ in vivo, as assayed by a modified mammalian twohybrid experiment. These data show that differential association of TEF-1 proteins with transcriptional co-activators may regulate the activity of TEF-1 family members.

Transcription factor TEAD2 is involved in neural tube closure

“…These transcription factors are defined by the presence of the 72 amino acid TEA DNA binding domain in their N-terminal half that is virtually identical among TEAD proteins and by the conspicuous homology among their C-terminal halves (Kaneko and DePamphilis, 1998). This conservation of structure is reflected in the ability of these proteins to bind the same transcriptional coactivator proteins (Mahoney et al, 2005;Vassilev et al, 2001) and to substitute for TEAD proteins in other organisms (Deshpande et al, 1997). Nevertheless, despite their structural similarities, the four mammalian TEAD proteins are not functionally redundant, because mice lacking TEAD1 fail to develop a proper heart and die between embryonic day 11 (E11) and E12 (Chen et al, 1994), whereas mice lacking TEAD4 arrest development before E3.5 (Yagi et al, 2007).…”

Yap1 is dispensable for self‐renewal but required for proper differentiation of mouse embryonic stem ( ES ) cells

“…We further investigated the activity of nuclear Yap1 using a synthetic Yap1-responsive luciferase (8xGTIIC) construct as previously designed for the measurement of Yap1 transcriptional activity in mechanical stress condition (Fig 3H) [15,19,31,32]. The luciferase construct contains repeated Yap1-Tead binding motifs (eight times) in front of the minimal cTNT promoter followed by a luciferase reporter gene [15,32,33].…”

Insights into transcription enhancer factor 1 (TEF-1) activity from the solution structure of the TEA domain

“…This sequence conservation is sufficient for functional conservation because TEF-1 substitutes for SD in wing development (24). The gene regulatory activities of TEF-1 are governed by interactions with protein cofactors (25)(26)(27). For instance, interactions of TEF-1 with Max and serum response factor (SRF) regulate the ␣-MHC gene and normal cardiac and smooth muscle development (28,29).…”