Atlas of Genetics and Cytogenetics

in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS

Gene Section Review

HUS1 (HUS1 checkpoint homolog (S. pombe)) Amrita Madabushi, Randall C Gunther, A-Lien Lu Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, 108 North Greene Street, Baltimore, Maryland 21201, USA (AM, RCG, ALL)

Published in Atlas Database: September 2010 Online updated version : http://AtlasGeneticsOncology.org/Genes/HUS1ID40899ch7p12.html DOI: 10.4267/2042/45024 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2011 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity HGNC (Hugo): HUS1 Description Location: 7p12.3 Amino acids: 280. Molecular weight: 31.7 kDa. Hus1 Note: NCBI accession: NM_004507.2; NP_004498.1. is a component of the 9-1-1 cell cycle checkpoint complex that plays a critical role in sensing DNA DNA/RNA damage and maintaining genomic stability. Description Expression Found in all tissues. 15464 bp; 8 exons. Transcription Localisation Nucleus and cytoplasm. In discrete nuclear foci upon The transcribed mRNA has 2143 bp and the coding DNA damage. region is 840 bp, encodes a 280 amino acids, 31691 Da protein.

Pseudogene None.

Figure 1. HUS1 adapted from NCBI database Homo sapiens 7, GrCh37 primary reference assembly with kilobases from the telomere of p-arm on bottom. The exons 1-8 are represented by boxes with transcribed and untranscribed sequences in pink and orange, respectively. The exon numbers are labeled on top. The grey arrowhead symbolizes the direction of transcription and the arrows show the ATG and the stop codons, respectively.

Atlas Genet Cytogenet Oncol Haematol. 2011; 15(5) 455 HUS1 (HUS1 checkpoint homolog (S. pombe)) Madabushi A, et al.

Function (MYH) has a strong preference to bind Hus1 (Chang and Lu, 2005; Shi et al., 2006). Hus1 along with and Rad1 forms the 9-1-1 complex Recent structural and functional analyses indicate that (Hang and Lieberman, 2000; St Onge et al., 1999; the Hus1 binding region of MYH adopts a stabilized Volkmer and Karnitz, 1999). Hus1 and Rad1 can also conformation projecting away from the catalytic form a stable 1:1 complex (Doré et al., 2009; Sohn and domain to form a docking scaffold for Hus1 and binds Cho, 2009; Xu et al., 2009). The 9-1-complex is loaded to Hus1 through electrostatic interaction (Luncsford et onto the DNA by a specific clamp loader (Rad17- al., 2010). RFC2-RFC3-RFC4-RFC5) in response to many The 9-1-1 complex is required to activate two different genotoxic stresses (alkylation, oxidation, checkpoint sensors-ATM (ataxia telangiectasia [AT] ultraviolet light radiation and ionizing radiation), and mutated protein) and ATR (ATM- and Rad3-related replication inhibitors (Bermudez et al., 2003; Ellison protein), which are phosphoinositol phosphate 3 and Stillman, 2003). The Rad1-Rad9 interface may be kinase-related kinases (PIKKs) (Zhou and Elledge, opened to encircle DNA (Doré et al., 2009; Sohn and 2000). The DNA-bound 9-1-1 complex facilitates Cho, 2009; Xu et al., 2009). ATM- or ATR-mediated phosphorylation of more than 700 including Chk1, Chk2, p53, and BRCA1 (Zhou and Elledge, 2000). Hus1 facilitated phosphorylation of Chk1 kinase is required for the ATR-dependent checkpoint; and regulates S-phase progression, G2/M arrest, and replication fork stabilization (Sancar et al., 2004; Sancar et al., 2004). However, Hus1 is not required for Chk2 phosphorylation in response to certain genotoxins (Weiss et al., 2003). Besides acting as a DNA damage sensor, the 9-1-1 complex plays an integral role in several DNA repair pathways including base excision repair (BER), mismatch repair (MMR), and nucleotide excision repair (NER) (see figure 2) (Helt et al., 2005). In the BER pathway, the 9-1-1 complex facilitates and interacts with several DNA glycosylases including MYH (Chang and Lu, 2005; Shi et al., 2006; Chang and Lu, 2005; Shi et al., 2006), 8-oxoG glycosylase (OGG1) (Park et al., 2009), NEIL1 (Guan et al., 2007a), and thymine DNA glycosylase (TDG) (Guan et Figure 2. hHus1 and hRad1 associate with each other prior to al., 2007b). The 9-1-1 complex also interacts with and forming the 9-1-1 complex with Rad9. In response to DNA stimulates other BER enzymes including APE1 damage and replication block, the 9-1-1 complex is loaded into (Gembka et al., 2007), POLbeta (Toueille et al., 2004), DNA by Rad17/RFC clamp loader and acts as checkpoint FEN1 (Friedrich-Heineken et al., 2005; Wang et al., sensor to activate ATM (ataxia telangiectasia [AT] mutated protein) or ATR (ATM- and Rad3-related protein) protein 2004a), RPA (Wu et al., 2005), and DNA ligase 1 kinase. The 9-1-1 complex interacts with and stimulates many (Smirnova et al., 2005; Wang et al., 2006a). Thus, the DNA repair enzymes involved in base excision repair (BER), 9-1-1 complex may provide a platform for the assembly nucleotide excision repair (NER), and mismatch repair (MMR). and function of the BER machinery (Balakrishnan et The damage recognition repair enzymes may serve as adaptors to signal DNA damage responses including enhanced DNA al., 2009; Lu et al., 2006). The 9-1-1 complex enhances repair, cell cycle arrest, and apoptosis. mismatch repair via direct interaction with mismatch The structure of the 9-1-1 complex (Doré et al., 2009; recognition proteins (MSH2/MSH3, MSH2/MSH6, and Sohn and Cho, 2009; Xu et al., 2009) is similar to the MLH1/PMS2) (Bai et al., 2010; He et al., 2008). Hus1 sliding clamp proliferating cell nuclear antigen protein interacts with MSH2/MSH3 and MSH2/MSH6, but not (PCNA) (Gulbis et al., 1996; Krishna et al., 1994). with MLH1/PMS2 (Bai et al., 2010; He et al., 2008). Hus1 interacts with Rad9 and Rad1 through its N In the NER pathway, interactions between terminal domain and C terminal domain, respectively. Saccharomyces cerevisiae Rad14 (hXPA homolog) and The structure and surface charge distribution of the the checkpoint proteins ScDdc1 (hRad9 homolog) and interdomain connecting loop (IDCL) of Hus1 differs ScMec3 (hHus1 homolog) have been demonstrated from those of other two subunits (Doré et al., 2009; (Giannattasio et al., 2004). Inactivation of NER by Sohn and Cho, 2009; Xu et al., 2009). The IDCL of knock down of XPA and XPC resulted in a decrease of Hus1 contains an N-terminal alpha helix and positive G1 phase cells that displayed Rad9 foci in response to charge cluster. These differences among Hus1, Rad9, UV light (Warmerdam et al., 2009). UV light-induced and Rad1 may contribute to different binding affinities Rad9 foci also colocalized with TopBP1 and gamma- to their partner proteins. For example, MutY homolog H2AX (Warmerdam et al., 2009).

Atlas Genet Cytogenet Oncol Haematol. 2011; 15(5) 456 HUS1 (HUS1 checkpoint homolog (S. pombe)) Madabushi A, et al.

Hus1 interacts with histone deacetylase HDAC1 (Cai et cells and impaired tissue regeneration (Yazinski et al., al., 2000). A novel pathway has been proposed that 2009). HDAC1 is involved in G(2)/M checkpoint control Oncogenesis through the interaction with the 9-1-1 complex. Single nucleotide polymorphism (SNP) analysis Jab1 physically associates with the 9-1-1 complex, supports the potential role of Hus1 in sporadic breast causes the translocation of the 9-1-1 complex from the cancer (Vega et al., 2009). SNPs in hHUS1 at nucleus to the cytoplasm, and mediates the rapid positions 47778020 (C) and degradation of the 9-1-1 complex (Huang et al., 2007). 47789957 (G) are statistically associated with breast Homology cancer development (Vega et al., 2009). hHus1 homologues from many eukaryotes are highly Ovarian tumors conserved. The following diagram shows the sequence Oncogenesis alignment of human Hus1 with Hus1 of fission yeast Hus1 expression levels correlate significantly with the Schizosaccharomyces pombe. The N-terminal domain clinicopathologic factors of bad prognosis of ovarian of Hus1 is structurally similar to the C-terminal tumors (de la Torre et al., 2008). domain. The structure of Hus1 (Doré et al., 2009; Sohn and Cho, 2009; Xu et al., 2009) is similar to those of Cancer therapy Rad9, Rad1, and PCNA (Gulbis et al., 1996; Krishna et Note al., 1994). The status of Hus1 can influence response to cancer therapy. Down-regulation of hHus1 by antisense RNA Mutations enhances the sensitivity of human lung carcinoma cells to cisplatin (a DNA cross-linker), presumably by Note enhancing apoptosis (Kinzel et al., 2002). Complete inactivation of the mouse Hus1 results in chromosomal instability, genotoxin hypersensitivity, Genomic stability and embryonic lethality. Note Inactivation of mouse or human Hus1 results in Implicated in impaired DNA damage signaling and severe spontaneous chromosomal instability (Weiss et al., Breast cancer 2002; Weiss et al., 2000; Kinzel et al., 2002). Note Drug sensitivity Hus1 inactivation in the mammary epithelium resulted in genome damage that induced apoptosis and led to Note depletion of Hus1-null cells from the mammary gland. Cells lacking Hus1 are hypersensitive to certain Dual inactivation of Hus1 and p53 in the mouse genotoxins including camptothecin (CPT), hydroxyurea mammary gland results in accumulation of damaged (HU), ultraviolet (UV), and ionizing radiation (IR) (Weiss et al., 2000; Wang et al., 2004b; Wang et al., 2006b).

Figure 3. Sequence alignment of the human Hus1 (hHus1) and S. pombe (SpHus1) with secondary structure motifs shown corresponding to the hHus1 based on hHus1 crystal structure (Xu et al., 2009). The arrows and coils represent the beta-sheets and alpha-helix, respectively. Hus1 contains 18 beta sheets and 5 alpha helices. Its N terminal and C terminal domains are connected by an interdomain connecting loop (IDCL, residues 134-155). The yellow shaded regions are the identical residues between hHus1 and SpHus1.

Atlas Genet Cytogenet Oncol Haematol. 2011; 15(5) 457 HUS1 (HUS1 checkpoint homolog (S. pombe)) Madabushi A, et al.

Loss of Hus1 sensitizes the cells to etoposide-induced Weiss RS, Matsuoka S, Elledge SJ, Leder P. Hus1 acts apoptosis by inducing Bim and Puma expressions and upstream of chk1 in a mammalian DNA damage response pathway. Curr Biol. 2002 Jan 8;12(1):73-7 releasing Rad9 into the cytosol (Levitt et al., 2005; Levitt et al., 2007; Meyerkord et al., 2008). The role of Bermudez VP, Lindsey-Boltz LA, Cesare AJ, Maniwa Y, Griffith JD, Hurwitz J, Sancar A. Loading of the human 9-1-1 Hus1 affecting the sensitivity of cells to IR-induced checkpoint complex onto DNA by the checkpoint clamp loader killing is independent of nonhomologous end-joining hRad17-replication factor C complex in vitro. Proc Natl Acad (NHEJ) but might be linked to homologous Sci U S A. 2003 Feb 18;100(4):1633-8 recombination repair (HRR) (Wang et al., 2006b). Ellison V, Stillman B. Biochemical characterization of DNA Development damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5' recessed DNA. PLoS Biol. Note 2003 Nov;1(2):E33 Targeted deletion of mouse Hus1 results in embryonic Weiss RS, Leder P, Vaziri C. Critical role for mouse Hus1 in an lethality (Weiss et al., 2000). S-phase DNA damage cell cycle checkpoint. Mol Cell Biol. 2003 Feb;23(3):791-803 Telomere maintenance Giannattasio M, Lazzaro F, Longhese MP, Plevani P, Muzi- Note Falconi M. Physical and functional interactions between Severe telomere shortening has been observed in both nucleotide excision repair and DNA damage checkpoint. Hus1-deficient mouse embryonic fibroblasts and EMBO J. 2004 Jan 28;23(2):429-38 thymocytes from conditional Hus1-knockout mice Sancar A, Lindsey-Boltz LA, Unsal-Kaçmaz K, Linn S. (Francia et al., 2006). Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem. 2004;73:39-85 HIV infection (AIDS) Toueille M, El-Andaloussi N, Frouin I, Freire R, Funk D, Note Shevelev I, Friedrich-Heineken E, Villani G, Hottiger MO, Hus1 is required for Human Immunodeficiency Virus Hübscher U. The human Rad9/Rad1/Hus1 damage sensor clamp interacts with DNA polymerase beta and increases its type 1 Vpr-mediated G2 cell cycle arrest (Zimmerman DNA substrate utilisation efficiency: implications for DNA et al., 2004). repair. Nucleic Acids Res. 2004;32(11):3316-24 Wang W, Brandt P, Rossi ML, Lindsey-Boltz L, Podust V, References Fanning E, Sancar A, Bambara RA. The human Rad9-Rad1- Hus1 checkpoint complex stimulates flap endonuclease 1. Krishna TS, Kong XP, Gary S, Burgers PM, Kuriyan J. Crystal Proc Natl Acad Sci U S A. 2004 Nov 30;101(48):16762-7 structure of the eukaryotic DNA polymerase processivity factor PCNA. Cell. 1994 Dec 30;79(7):1233-43 Wang X, Guan J, Hu B, Weiss RS, Iliakis G, Wang Y. Involvement of Hus1 in the chain elongation step of DNA Gulbis JM, Kelman Z, Hurwitz J, O'Donnell M, Kuriyan J. replication after exposure to camptothecin or ionizing radiation. Structure of the C-terminal region of p21(WAF1/CIP1) Nucleic Acids Res. 2004;32(2):767-75 complexed with human PCNA. Cell. 1996 Oct 18;87(2):297- 306 Zimmerman ES, Chen J, Andersen JL, Ardon O, Dehart JL, Blackett J, Choudhary SK, Camerini D, Nghiem P, Planelles V. St Onge RP, Udell CM, Casselman R, Davey S. The human Human immunodeficiency virus type 1 Vpr-mediated G2 arrest G2 checkpoint control protein hRAD9 is a nuclear requires Rad17 and Hus1 and induces nuclear BRCA1 and phosphoprotein that forms complexes with hRAD1 and hHUS1. gamma-H2AX focus formation. Mol Cell Biol. 2004 Mol Biol Cell. 1999 Jun;10(6):1985-95 Nov;24(21):9286-94 Volkmer E, Karnitz LM. Human homologs of Chang DY, Lu AL. Interaction of checkpoint proteins Schizosaccharomyces pombe rad1, hus1, and rad9 form a Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY DNA damage-responsive protein complex. J Biol Chem. 1999 homolog in fission yeast, Schizosaccharomyces pombe. J Biol Jan 8;274(2):567-70 Chem. 2005 Jan 7;280(1):408-17 Cai RL, Yan-Neale Y, Cueto MA, Xu H, Cohen D. HDAC1, a Friedrich-Heineken E, Toueille M, Tännler B, Bürki C, Ferrari histone deacetylase, forms a complex with Hus1 and Rad9, E, Hottiger MO, Hübscher U. The two DNA clamps two G2/M checkpoint Rad proteins. J Biol Chem. 2000 Sep Rad9/Rad1/Hus1 complex and proliferating cell nuclear 8;275(36):27909-16 antigen differentially regulate flap endonuclease 1 activity. J Hang H, Lieberman HB. Physical interactions among human Mol Biol. 2005 Nov 11;353(5):980-9 checkpoint control proteins HUS1p, RAD1p, and RAD9p, and Helt CE, Wang W, Keng PC, Bambara RA. Evidence that DNA implications for the regulation of cell cycle progression. damage detection machinery participates in DNA repair. Cell Genomics. 2000 Apr 1;65(1):24-33 Cycle. 2005 Apr;4(4):529-32 Weiss RS, Enoch T, Leder P. Inactivation of mouse Hus1 Levitt PS, Liu H, Manning C, Weiss RS. Conditional results in genomic instability and impaired responses to inactivation of the mouse Hus1 cell cycle checkpoint gene. genotoxic stress. Dev. 2000 Aug 1;14(15):1886-98 Genomics. 2005 Aug;86(2):212-24 Zhou BB, Elledge SJ. The DNA damage response: putting Smirnova E, Toueille M, Markkanen E, Hübscher U. The checkpoints in perspective. Nature. 2000 Nov human checkpoint sensor and alternative DNA clamp Rad9- 23;408(6811):433-9 Rad1-Hus1 modulates the activity of DNA ligase I, a Kinzel B, Hall J, Natt F, Weiler J, Cohen D. Downregulation of component of the long-patch base excision repair machinery. Hus1 by antisense oligonucleotides enhances the sensitivity of Biochem J. 2005 Jul 1;389(Pt 1):13-7 human lung carcinoma cells to cisplatin. Cancer. 2002 Mar 15;94(6):1808-14

Atlas Genet Cytogenet Oncol Haematol. 2011; 15(5) 458 HUS1 (HUS1 checkpoint homolog (S. pombe)) Madabushi A, et al.

Wu X, Shell SM, Zou Y. Interaction and colocalization of Meyerkord CL, Takahashi Y, Araya R, Takada N, Weiss RS, Rad9/Rad1/Hus1 checkpoint complex with replication protein A Wang HG. Loss of Hus1 sensitizes cells to etoposide-induced in human cells. Oncogene. 2005 Jul 7;24(29):4728-35 apoptosis by regulating BH3-only proteins. Oncogene. 2008 Dec 11;27(58):7248-59 Francia S, Weiss RS, Hande MP, Freire R, d'Adda di Fagagna F. Telomere and telomerase modulation by the mammalian Balakrishnan L, Brandt PD, Lindsey-Boltz LA, Sancar A, Rad9/Rad1/Hus1 DNA-damage-checkpoint complex. Curr Biol. Bambara RA. Long patch base excision repair proceeds via 2006 Aug 8;16(15):1551-8 coordinated stimulation of the multienzyme DNA repair complex. J Biol Chem. 2009 May 29;284(22):15158-72 Lu AL, Bai H, Shi G, Chang DY. MutY and MutY homologs (MYH) in genome maintenance. Front Biosci. 2006 Sep Doré AS, Kilkenny ML, Rzechorzek NJ, Pearl LH. Crystal 1;11:3062-80 structure of the rad9-rad1-hus1 DNA damage checkpoint complex--implications for clamp loading and regulation. Mol Shi G, Chang DY, Cheng CC, Guan X, Venclovas C, Lu AL. Cell. 2009 Jun 26;34(6):735-45 Physical and functional interactions between MutY glycosylase homologue (MYH) and checkpoint proteins Rad9-Rad1-Hus1. Park MJ, Park JH, Hahm SH, Ko SI, Lee YR, Chung JH, Sohn Biochem J. 2006 Nov 15;400(1):53-62 SY, Cho Y, Kang LW, Han YS. Repair activities of human 8- oxoguanine DNA glycosylase are stimulated by the interaction Wang W, Lindsey-Boltz LA, Sancar A, Bambara RA. with human checkpoint sensor Rad9-Rad1-Hus1 complex. Mechanism of stimulation of human DNA ligase I by the Rad9- DNA Repair (Amst). 2009 Oct 2;8(10):1190-200 rad1-Hus1 checkpoint complex. J Biol Chem. 2006 Jul 28;281(30):20865-72 Sohn SY, Cho Y. Crystal structure of the human rad9-hus1- rad1 clamp. J Mol Biol. 2009 Jul 17;390(3):490-502 Wang X, Hu B, Weiss RS, Wang Y. The effect of Hus1 on ionizing radiation sensitivity is associated with homologous Vega A, Salas A, Milne RL, Carracedo B, Ribas G, Ruibal A, recombination repair but is independent of nonhomologous de León AC, González-Hernández A, Benítez J, Carracedo A. end-joining. Oncogene. 2006 Mar 23;25(13):1980-3 Evaluating new candidate SNPs as low penetrance risk factors in sporadic breast cancer: a two-stage Spanish case-control Gembka A, Toueille M, Smirnova E, Poltz R, Ferrari E, Villani study. Gynecol Oncol. 2009 Jan;112(1):210-4 G, Hübscher U. The checkpoint clamp, Rad9-Rad1-Hus1 complex, preferentially stimulates the activity of Warmerdam DO, Freire R, Kanaar R, Smits VA. Cell cycle- apurinic/apyrimidinic endonuclease 1 and DNA polymerase dependent processing of DNA lesions controls localization of beta in long patch base excision repair. Nucleic Acids Res. Rad9 to sites of genotoxic stress. Cell Cycle. 2009 Jun 2007;35(8):2596-608 1;8(11):1765-74 Guan X, Bai H, Shi G, Theriot CA, Hazra TK, Mitra S, Lu AL. Xu M, Bai L, Gong Y, Xie W, Hang H, Jiang T. Structure and The human checkpoint sensor Rad9-Rad1-Hus1 interacts with functional implications of the human rad9-hus1-rad1 cell cycle and stimulates NEIL1 glycosylase. Nucleic Acids Res. checkpoint complex. J Biol Chem. 2009 Jul 31;284(31):20457- 2007;35(8):2463-72 61 Guan X, Madabushi A, Chang DY, Fitzgerald ME, Shi G, Yazinski SA, Westcott PM, Ong K, Pinkas J, Peters RM, Weiss Drohat AC, Lu AL. The human checkpoint sensor Rad9-Rad1- RS. Dual inactivation of Hus1 and p53 in the mouse mammary Hus1 interacts with and stimulates DNA repair enzyme TDG gland results in accumulation of damaged cells and impaired glycosylase. Nucleic Acids Res. 2007;35(18):6207-18 tissue regeneration. Proc Natl Acad Sci U S A. 2009 Dec 15;106(50):21282-7 Huang J, Yuan H, Lu C, Liu X, Cao X, Wan M. Jab1 mediates protein degradation of the Rad9-Rad1-Hus1 checkpoint Bai H, Madabushi A, Guan X, Lu AL. Interaction between complex. J Mol Biol. 2007 Aug 10;371(2):514-27 human mismatch repair recognition proteins and checkpoint sensor Rad9-Rad1-Hus1. DNA Repair (Amst). 2010 May Levitt PS, Zhu M, Cassano A, Yazinski SA, Liu H, Darfler J, 4;9(5):478-87 Peters RM, Weiss RS. Genome maintenance defects in cultured cells and mice following partial inactivation of the Luncsford PJ, Chang DY, Shi G, Bernstein J, Madabushi A, essential cell cycle checkpoint gene Hus1. Mol Cell Biol. 2007 Patterson DN, Lu AL, Toth EA. A structural hinge in eukaryotic Mar;27(6):2189-201 MutY homologues mediates catalytic activity and Rad9-Rad1- Hus1 checkpoint complex interactions. J Mol Biol. 2010 Oct de la Torre J, Gil-Moreno A, García A, Rojo F, Xercavins J, 29;403(3):351-70 Salido E, Freire R. Expression of DNA damage checkpoint protein Hus1 in epithelial ovarian tumors correlates with This article should be referenced as such: prognostic markers. Int J Gynecol Pathol. 2008 Jan;27(1):24- 32 Madabushi A, Gunther RC, Lu AL. HUS1 (HUS1 checkpoint homolog (S. pombe)). Atlas Genet Cytogenet Oncol Haematol. He W, Zhao Y, Zhang C, An L, Hu Z, Liu Y, Han L, Bi L, Xie Z, 2011; 15(5):455-459. Xue P, Yang F, Hang H. Rad9 plays an important role in DNA mismatch repair through physical interaction with MLH1. Nucleic Acids Res. 2008 Nov;36(20):6406-17

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