II. Structure and specificity of the interaction between the FHA2 domain of rad53 and phosphotyrosyl peptides

Wang, P; Byeon, I.-J.L.; Liao, Hua; Beebe, Kirk; Yongkiettrakul, Suganya; Pei, Dehua; Tsai, Ming‐Daw

doi:10.1006/jmbi.2000.4095

Cited by 52 publications

(56 citation statements)

References 34 publications

(53 reference statements)

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“…Three phosphopeptides were used to examine the affinities of the four FHA mutants for phosphopeptides: SLEV(pT)EADATFVQ (peptide I), EVEL(pT)QELP (peptide II), and EDI(pY)YLD (peptide III, a phosphotyrosine peptide). All three phosphopeptides correspond to partial sequences of Rad9, a potential binding partner of the FHA1 and FHA2 domains (7,18,19). The structures of the FHA1-peptide I, FHA2-peptide II, and FHA2-peptide III complexes have all been determined by NMR in our laboratory, and their binding properties have been characterized (7,(18)(19)(20).…”

Section: Resultsmentioning

confidence: 99%

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,

2004

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On the basis of the results from our laboratory and others, we recently suggested that the ligand specificity of forkhead-associated (FHA) domains is controlled by variations in three major factors: (i) residues interacting with pThr, (ii) residues recognizing the +1 to +3 residues from pThr, and (iii) an extended binding surface. While the first factor has been well established by several solution and crystal structures of FHA-phosphopeptide complexes, the structural bases of the second and third factors are not well understood and are likely to vary greatly between different FHA domains. In this work, we proposed and tested the hypothesis that nonconserved residues G133 and G135 of FHA1 and I681 and D683 of FHA2, located outside of the core FHA region of yeast Rad53 FHA domains, contribute to the specific recognition of the +3 position of different phosphopeptides. By rational mutagenesis of these residues, the specificity of FHA1 has been changed from predominantly pTXXD to be equally acceptable for pTXXD, pTXXL, and pYXL, which are similar to the specificities of the FHA2 domain of Rad53. Conversely, the +3 position specificity of FHA2 has been engineered to be more like FHA1 with the I681A mutation. These results were based on library screening as well as binding analyses of specific phosphopeptides. Furthermore, results of structural analyses by NMR indicate that some of these residues are also important for the structural integrity of the loops.Recently, a phosphorylation-dependent recognition domain, known as the forkhead-associated (FHA) 1 domain, has been shown to mediate protein-protein interactions in many cellular processes of different species (1-4). Although the FHA domain has been found in more than 200 different proteins, the tertiary structure has only been obtained for yeast Rad53 (which consists of two FHA domains, FHA1 at the N-terminus and FHA2 at the C-terminus), human Chk2, human Chfr, human Ki67, and plant kinase-associated protein phosphatase (KAPP) (5-11). While the level of sequence homology of FHA domains is relatively low (∼20%), the tertiary structures reveal a remarkably similar folding, consisting of -strands linked by several loops to form two large twisted antiparallel -sheets folded into a -sandwich. The biological functions of FHA domains are very diverse. Therefore, the structure-function relationship of FHA domains and the structural basis of interactions between FHA domains and their binding proteins have been subjects of extensive research in many laboratories. The biological binding protein and the actual binding site have been unequivocally identified in only very few cases (12-16). Nevertheless, a wealth of information about the ligand specificity of FHA domains has been obtained by use of short phosphopeptides and phosphopeptide libraries consisting of 6-15 residues (5-7, 9, 17-20). These studies with short phosphopeptides showed that, while the Rad53 FHA1 domain specifically recognizes the phosphothreonine peptides containing Asp at the +3 position (pTXXD),...

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Section: Resultsmentioning

confidence: 99%

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,

2004

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“…Extracts from these cells were immunoblotted for Chfr after treatment with lambda phosphatase (PPase) or buffer, as described in Materials and methods ubiquitin-conjugating enzymes (Joazeiro et al, 1999;Levkowitz et al, 1999;Lorick et al, 1999;Ciechanover et al, 2000;Zheng et al, 2000). Second, Chfr has an FHA domain, a type of domain involved in proteinprotein interactions (Durocher et al, 1999(Durocher et al, , 2000Liao et al, 1999;Wang et al, 2000;Stavridi et al, 2002). Ubiquitin ligases typically contain protein-protein interaction domains to recognize their substrates.…”

Section: Discussionmentioning

confidence: 99%

“…At its N-terminus, Chfr has a forkhead-associated (FHA) domain. FHA domains from other proteins bind phosphopeptides (Durocher et al, 1999(Durocher et al, , 2000Liao et al, 1999;Wang et al, 2000). The three-dimensional structure of the Chfr FHA domain bound to tungstate, an analog of phosphate, suggests that Chfr also recognizes phosphorylated proteins (Stavridi et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63-linked polyubiquitin chains

et al. 2003

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“…These are the only FHA domains whose tertiary structures have been determined in the free form. 5,8,9 Despite relatively low sequence homology, they are structurally similar, consisting mainly of antiparallel b-sheets that fold into a twisted bsandwich. Signi®cant differences, however, are noticeable in some of the elements (turns, loops, and a-helix) that connect 11 structurally equivalent b-strands.…”

Section: Introductionmentioning

confidence: 99%

“…The FHA2 domain has been demonstrated to bind both pY-peptides 5,9 and pTpeptides. 10,11 For FHA1, a consensus ligand motif of pTXXD has been identi®ed, where Asp is required at the 3 position and Ala/Ile is preferred at the 2 position for high-af®nity recognition.…”

Section: Introductionmentioning

confidence: 99%

Solution structures of two FHA1-phosphothreonine peptide complexes provide insight into the structural basis of the ligand specificity of FHA1 from yeast Rad53 1 1Edited by M. F. Summers

Yuan

Yongkiettrakul

Byeon

et al. 2001

Journal of Molecular Biology

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II. Structure and specificity of the interaction between the FHA2 domain of rad53 and phosphotyrosyl peptides

Cited by 52 publications

References 34 publications

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,

The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63-linked polyubiquitin chains

Solution structures of two FHA1-phosphothreonine peptide complexes provide insight into the structural basis of the ligand specificity of FHA1 from yeast Rad53 1 1Edited by M. F. Summers

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II. Structure and specificity of the interaction between the FHA2 domain of rad53 and phosphotyrosyl peptides

Cited by 52 publications

References 34 publications

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues,

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues,

The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63-linked polyubiquitin chains

Solution structures of two FHA1-phosphothreonine peptide complexes provide insight into the structural basis of the ligand specificity of FHA1 from yeast Rad53 1 1Edited by M. F. Summers

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The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,

The Ligand Specificity of Yeast Rad53 FHA Domains at the +3 Position Is Determined by Nonconserved Residues^,