Download MendeleyStructure of the FHA1 domain of yeast Rad53 and identification of binding sites for both FHA1 and its target protein Rad9
Liao, H., Yuan, C., Su, M.I., Yongkiettrakul, S., Qin, D., Li, H., Byeon, I.J., Pei, D., Tsai, M.D.(2000) J Mol Biol 304: 941-951
- PubMed: 11124038
- DOI: https://doi.org/10.1006/jmbi.2000.4291
- Primary Citation of Related Structures:
1G3G - PubMed Abstract:
Forkhead-associated (FHA) domains have been shown to recognize both pThr and pTyr-peptides. The solution structures of the FHA2 domain of Rad53 from Saccharomyces cerevisiae, and its complex with a pTyr peptide, have been reported recently. We now report the solution structure of the other FHA domain of Rad53, FHA1 (residues 14-164), and identification of binding sites of FHA1 and its target protein Rad9. The FHA1 structure consists of 11 beta-strands, which form two large twisted anti-parallel beta-sheets folding into a beta-sandwich. Three short alpha-helices were also identified. The beta-strands are linked by several loops and turns. These structural features of free FHA1 are similar to those of free FHA2, but there are significant differences in the loops. Screening of a peptide library [XXX(pT)XXX] against FHA1 revealed an absolute requirement for Asp at the +3 position and a preference for Ala at the +2 position. These two criteria are met by a pThr motif (192)TEAD(195) in Rad9. Surface plasmon resonance analysis showed that a pThr peptide containing this motif, (188)SLEV(pT)EADATFVQ(200) from Rad9, binds to FHA1 with a K(d) value of 0.36 microM. Other peptides containing pTXXD sequences also bound to FHA1, but less tightly (K(d)=4-70 microM). These results suggest that Thr192 of Rad9 is the likely phosphorylation site recognized by the FHA1 domain of Rad53. The tight-binding peptide was then used to identify residues of FHA1 involved in the interaction with the pThr peptide. The results are compared with the interactions between the FHA2 domain and a pTyr peptide derived from Rad9 reported previously.
Organizational Affiliation:
Departments of Chemistry and Biochemistry, The Ohio State Biochemistry Program, and Campus Chemical Instrument Center, The Ohio State University, Columbus, OH 43210, USA.
