The FHA Domain Is a Modular Phosphopeptide Recognition Motif

Durocher, Daniel; Henckel, Julia; Fersht, Alan R.; Jackson, Stephen

doi:10.1016/s1097-2765(00)80340-8

Cited by 370 publications

(343 citation statements)

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“…This behavior is consistent with the studies of pTyr peptide recognition. The pT+3 position of peptide ligands is the most important determinant of their affinity for FHA domains (3,79). The non-conserved residues that divergent FHA domains place around the pT+3 site appear to be important for affinity (77).…”

Section: Rigidity and Role In Affinity Of Residues At The Pt+3-site Amentioning

confidence: 99%

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,

et al. 2005

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A net increase in the backbone rigidity of the kinase-interacting FHA domain (KI-FHA) from the Arabidopsis receptor kinase-associated protein phosphatase (KAPP) accompanies the binding of a phosphoThr peptide from its CLV1 receptor-like kinase partner, according to 15 N NMR relaxation at 11.7 and 14.1 T. All of the loops of free KI-FHA display evidence of nsec-scale motions. Many of these same residues have residual dipolar couplings that deviate from structural predictions. Binding of the CLV1 pT868 peptide seems to reduce nsec-scale fluctuations of all loops, including half of the residues of recognition loops. Residues important for affinity are found to be rigid, i.e. conserved residues and residues of the subsite for the key pT+3 peptide position. -This behavior parallels SH2 and PTB domain recognition of pTyr peptides. PhosphoThr peptide binding increases KI-FHA backbone rigidity (S 2 ) of three recognition loops, a loop nearby, seven strands from the β-sandwich, and a distal loop. Compensating the trend of increased rigidity, binding enhances fast mobility at a few sites in four loops on the periphery of the recognition surface and in two loops on the far side of the β-sandwich. Line broadening evidence of µsec to msec-scale fluctuations occurs across the six-stranded β-sheet and nearby edges of the β-sandwich; this forms a network connected by packing of interior side chains and H-bonding. A patch of the slowly fluctuating residues coincides with the site of segment-swapped dimerization in crystals of the FHA domain of human Chfr. Phosphopeptide binding introduces µsec to msec-scale fluctuations to more residues of the long 8/9 *To whom correspondence should be addressed, E-mail: vandorens@missouri.edu, Phone: 1 (573) 882-5113, FAX: 1 (573) 884-4812. † These authors contributed equally to this work. ‡ Current address: Department of Chemistry, 225 Prospect St., Yale University, New Haven, CT 06520The NMR relaxation data and their model-free interpretation are available for free and bound KI-FHA under BMRB accession codes 5841 and 6474 at www.bmrb.wisc.edu. NIH Public Access Author ManuscriptBiochemistry. Author manuscript; available in PMC 2010 January 29. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript recognition loop of KI-FHA. The rigidity of this FHA domain appears to couple as a whole to pThr peptide binding. KeywordsFHA domain; phosphoThr-binding module; protein-protein interactions; NMR relaxation; relaxation dispersion; dynamics; residual dipolar couplings FHA domains bind phosphothreonine-and phosphoserine-containing partners in diverse eukaryotic signaling pathways that include DNA damage repair, cell proliferation, pre-mRNA splicing, forkhead transcription factors, Ring-finger proteins, and kinesins (1). Several highresolution structures of FHA domains have appeared (2-8), but no detailed studies of their dynamics. Flexibility often appears to correlate with binding events, including affinity of protein modules for peptides (9-11). Residue-specific effects on...

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Section: Rigidity and Role In Affinity Of Residues At The Pt+3-site Amentioning

confidence: 99%

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,

et al. 2005

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“…The four Calymmin prodomains exhibit different degrees of homology to portions of the amino acid sequence of some polypeptides from the databases with unknown functions. These peptides included the Drosophila melanogaster cg6639, a serine protease-like peptide, and cg11581 proteins (Adams et al, 2000), and the Mycobacterium tuberculosis hypothetical 56.9-kDa protein cy10h4.20c (Cole et al, 1998), and the M. leprae hypothetical 49.8-kDa protein (Eiglmeier, et al, 1993), both of which also contain forkhead-associated domains (Durocher et al, 1999) at their C terminus ( Fig. 2A).…”

Section: Isolation Of Calymmin Full-length Cdna and Amino Acid Sequenmentioning

confidence: 99%

Molecular characterization of Calymmin, a novel notochord sheath‐associated extracellular matrix protein in the zebrafish embryo

Cerdà

Gründ

Franke

et al. 2002

Developmental Dynamics

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During the screening of a zebrafish postsomitogenesis embryo cDNA library, we have identified a cDNA corresponding to a novel type of protein localized to the notochordal sheath-associated extracellular matrix (ECM) of the embryo. The 4.049-kb mRNA encodes a predicted polypeptide of 1,207 amino acids (122 kDa, pI 10.50) with a potential signal peptide of 20 amino acids. After the signal peptide, the mature protein consists of 1,187 amino acids (119 kDa, pI 10.46), for which the name "Calymmin" (from Greek ␣ ␣, to envelop, to cover) is proposed. The Calymmin mRNA is highly and transiently expressed by the notochord cells of the embryo from the 10-to 12-somite stage to the pharyngula period (13 and 24 hours postfertilization, respectively), and light and electron microscopical immunolocalization analysis revealed that the protein was specifically localized within a granular and filamentous layer of the ECM compartment surrounding the notochord. In zebrafish no tail mutants (ntl tc41 ), in which the notochord precursor cells are present but fail to differentiate, the Calymmin protein was not detected, confirming the notochord origin of Calymmin. These results indicate that Calymmin is a novel constitutive protein of the ECM compartment associated to the perinotochordal sheath in the zebrafish embryo, which is specifically expressed by the differentiating notochord cells.

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“…The N terminus includes a forkhead-associated (FHA) domain (a.a. 20-108) and two BRCA1 C terminus (BRCT) domains (BRCT1, a.a. BRCT2,. The FHA domain has been shown to be a proteinprotein interaction motif, and the BRCT domain is also thought to be an interaction domain; these motifs bind to phosphorylated regions of specific proteins (Durocher et al, 1999). Indeed, we previously reported an interaction between the FHA and BRCT1 domains of NBS1 and phosphorylated histone H2AX that was responsible for recruitment of NBS1 to the vicinity of DSB sites (Kobayashi et al, 2002).…”

Section: Introductionmentioning

confidence: 93%

Chromatin modification and NBS1: their relationship in DNA double-strand break repair

2015

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The importance of chromatin modification, including histone modification and chromatin remodeling, for DNA double-strand break (DSB) repair, as well as transcription and replication, has been elucidated. Phosphorylation of H2AX to γ-H2AX is one of the first responses following DSB detection, and this histone modification is important for the DSB damage response by triggering several events, including the accumulation of DNA damage response-related proteins and subsequent homologous recombination (HR) repair. The roles of other histone modifications such as acetylation, methylation and ubiquitination have also been recently clarified, particularly in the context of HR repair. NBS1 is a multifunctional protein that is involved in various DNA damage responses. Its recently identified binding partner RNF20 is an E3 ubiquitin ligase that facilitates the monoubiquitination of histone H2B, a process that is crucial for recruitment of the chromatin remodeler SNF2h to DSB damage sites. Evidence suggests that SNF2h functions in HR repair, probably through regulation of end-resection. Moreover, several recent reports have indicated that SNF2h can function in HR repair pathways as a histone remodeler and that other known histone remodelers can also participate in DSB damage responses. On the other hand, information about the roles of such chromatin modifications and NBS1 in non-homologous end joining (NHEJ) repair of DSBs and stalled fork-related damage responses is very limited; therefore, these aspects and processes need to be further studied to advance our understanding of the mechanisms and molecular players involved.

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The FHA Domain Is a Modular Phosphopeptide Recognition Motif

Cited by 370 publications

References 33 publications

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,

Molecular characterization of Calymmin, a novel notochord sheath‐associated extracellular matrix protein in the zebrafish embryo

Chromatin modification and NBS1: their relationship in DNA double-strand break repair

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The FHA Domain Is a Modular Phosphopeptide Recognition Motif

Cited by 370 publications

References 33 publications

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase,

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase,

Molecular characterization of Calymmin, a novel notochord sheath‐associated extracellular matrix protein in the zebrafish embryo

Chromatin modification and NBS1: their relationship in DNA double-strand break repair

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Product

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PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,

PhosphoThr Peptide Binding Globally Rigidifies Much of the FHA Domain from Arabidopsis Receptor Kinase-Associated Protein Phosphatase^,