The FHA Domain

Durocher, Daniel

doi:10.1002/3527603611.ch7

Cited by 129 publications

(185 citation statements)

References 61 publications

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“…The two FHA domains of ScRad53p are known to interact with different upstream and downstream proteins, resulting in different responses to different genotoxic insults (Durocher et al, 1999;Durocher and Jackson, 2002;Emili et al, 2001;Duncker et al, 2002;Tsai, 2002;Leroy et al, 2003;Schwartz et al, 2003). Thus, we asked whether the FHA domains of CaRad53p are required for the filamentous growth induced by genotoxins.…”

Section: Role Of Fha Domains In Rad53p-mediated Cellular Functionsmentioning

confidence: 99%

“…The two FHA domains of ScRad53p are known to mediate associations with different partners (Durocher and Jackson, 2002;Tsai, 2002;Schwartz et al, 2003), and FHA-domain mutations can alter their substrate specificity (Durocher et al, 2000;. We reasoned that if CaRad53p promotes filamentous growth through FHA-domain-mediated interactions with other proteins, it might be possible to find FHA-domain mutations that block or compromise the genotoxicstress-induced filamentous growth without affecting other Rad53p-controlled responses.…”

Section: Genetic Separation Of Rad53p-mediated Responses To Genotoxicmentioning

confidence: 99%

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Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

Shi

Wang

Zheng

et al. 2007

MBoC

119

198

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The polymorphic fungus Candida albicans switches from yeast to filamentous growth in response to a range of genotoxic insults, including inhibition of DNA synthesis by hydroxyurea (HU) or aphidicolin (AC), depletion of the ribonucleotidereductase subunit Rnr2p, and DNA damage induced by methylmethane sulfonate (MMS) or UV light (UV). Deleting RAD53, which encodes a downstream effector kinase for both the DNA-replication and DNA-damage checkpoint pathways, completely abolished the filamentous growth caused by all the genotoxins tested. Deleting RAD9, which encodes a signal transducer of the DNA-damage checkpoint, specifically blocked the filamentous growth induced by MMS or UV but not that induced by HU or AC. Deleting MRC1, the counterpart of RAD9 in the DNA-replication checkpoint, impaired DNA synthesis and caused cell elongation even in the absence of external genotoxic insults. Together, the results indicate that the DNA-replication/damage checkpoints are critically required for the induction of filamentous growth by genotoxic stress. In addition, either of two mutations in the FHA1 domain of Rad53p, G65A, and N104A, nearly completely blocked the filamentous-growth response but had no significant deleterious effect on cell-cycle arrest. These results suggest that the FHA domain, known for its ability to bind phosphopeptides, has an important role in mediating genotoxic-stress-induced filamentous growth and that such growth is a specific, Rad53p-regulated cellular response in C. albicans. INTRODUCTIONThe polymorphic fungus Candida albicans has proven to be a very useful model for the study of cell morphogenesis (Odds, 1994;Sudbery et al., 2004;Wightman et al., 2004;Zheng et al., 2004). This organism can switch between several morphological forms including yeast, pseudohyphae, and hyphae (Berman and Sudbery, 2002;Sudbery et al., 2004). The yeast-hypha transition has attracted much attention because of its established importance for infection and virulence (Csank et al., 1997;Lo et al., 1997;Gale et al., 1998;Gow et al., 2002;Zheng et al., 2004). Several signal-transduction pathways are known to play roles in growth-form selection (Liu, 2001;Berman and Sudbery, 2002). The cAMPprotein kinase A (PKA) pathway has a key role, because blocking it abolishes true hyphal growth under most experimental conditions (Lo et al., 1997;Stoldt et al., 1997;Rocha et al., 2001). Another important pathway contains a MAPkinase cascade that seems to have a more important role in regulating pseudohyphal than true hyphal growth (Liu et al., 1994). The two pathways control the transcription factors Efg1p and Cph1p, respectively, which activate the expression of hypha-specific genes for a suite of infection-related functions (Lane et al., 2001).A diverse range of environmental stimuli can induce C. albicans hyphal or pseudohyphal growth, such as serum, neutral pH, appropriate temperature, certain amino acids and sugars, and some synthetic growth media (Ernst, 2000;Sudbery et al., 2004). These conditions more-or-less mimic certain host c...

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Section: Role Of Fha Domains In Rad53p-mediated Cellular Functionsmentioning

confidence: 99%

Section: Genetic Separation Of Rad53p-mediated Responses To Genotoxicmentioning

confidence: 99%

Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

Shi

Wang

Zheng

et al. 2007

MBoC

119

198

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“…RDCs were obtained by comparing coupled 15 N IPAP-HSQC spectra of KI-FHA in the absence of orienting media against spectra of KI-FHA weakly oriented using the phage or PEG/hexanol media. 1 D NH values of successive sets of five neighboring residues were fitted by singular value decomposition (SVD) to the four lowest energy structural models of KI-FHA (PDB code 1MZK) to obtain alignment tensor parameters as described (44). The application here is novel in using a group of NMR structures rather than an X-ray structure.…”

Section: Residual Dipolar Coupling Analysismentioning

confidence: 99%

“…β8 and the C-terminus also have residues with deviating 1 D NH RDCs (Figure 3b). A majority of the sites of 1 We also used the method of ref (44) to fit successive quintets of 1 D NH RDCs to the atomic coordinates of their respective fragments of the KI-FHA structural model. Deviations of molecular segments' alignment tensors from the overall protein alignment may result from dynamic or static deviations (43).…”

Section: Fluctuations In the Loops Of Free Ki-fha Are Corroborated Bymentioning

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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“…FHA domains were first identified in forkhead-type transcription factors, but are now known to exist in various proteins (Hofmann and Bucher, 1995). Many FHA-containing proteins play important roles in the regulation of DNA repair, the cell cycle and apoptosis (Durocher and Jackson, 2002). More than 200 other FHA-containing proteins have also been identified; however, the functions of many of these proteins remain unknown.…”

Section: Introductionmentioning

confidence: 99%

CHFR, a potential tumor suppressor, downregulates interleukin-8 through the inhibition of NF-κB

et al. 2009

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The mitotic checkpoint gene CHFR (checkpoint with forkhead and ring finger domains) is silenced in various human cancers by promoter hypermethylation, suggesting that CHFR is a tumor suppressor. Here, we show that CHFR functions as a negative regulator of the nuclear factor-jB (NF-jB) pathway. Expression of CHFR inhibited NF-jB reporter activity, whereas knockdown of CHFR activated reporter activity. These activities are independent of its RING finger domain. Furthermore, we found that CHFR physically interacts with p65 in cells. Electrophoretic mobility shift assays (EMSAs) and ELISA-based NF-jB-binding assays showed that CHFR negatively regulated transcriptional activity of p65. In addition, our data show that interleukin (IL)-8 is significantly downregulated by CHFR, and that the migration of human endothelial cells is suppressed in culture medium conditioned from CHFR-expressing cancer cells. Using a xenograft model, we show that neovascularization is suppressed by adenovirus-mediated transfer of CHFR. These results indicate that expression of CHFR markedly reduces the expression of IL-8 through the inhibition of NF-jB. As the NF-jB signaling pathway plays a critical role in the development and progression of cancer, our findings show the functional relationship between epigenetic alteration and inflammation/angiogenesis in human cancer cells, thereby showing several potential targets for therapeutic intervention.

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The FHA Domain

Cited by 129 publications

References 61 publications

Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

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

CHFR, a potential tumor suppressor, downregulates interleukin-8 through the inhibition of NF-κB

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The FHA Domain

Cited by 129 publications

References 61 publications

Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

Critical Role of DNA Checkpoints in Mediating Genotoxic-Stress–induced Filamentous Growth inCandida albicans

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

CHFR, a potential tumor suppressor, downregulates interleukin-8 through the inhibition of NF-κB

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