Molecular cloning and tissue-specific expression ofMrad9, a murine orthologue of theSchizosaccharomyces pombe rad9+ checkpoint control gene

Hang, Haiying; Rauth, Sarah J.; Hopkins, Kevin M.; Davey, Scott; Lieberman, Howard B.

doi:10.1002/(sici)1097-4652(199811)177:2<241::aid-jcp6>3.0.co;2-n

Cited by 15 publications

(17 citation statements)

References 31 publications

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“…Although this function for Rad9 was never tested, the lack of proapoptosis activity for the protein is unlikely since such a function was demonstrated for the human and fission yeast orthologs. In addition, the encoded mouse protein is structurally very similar to the RAD9 gene product (11), and it also contains a BH3-like domain characteristic of pro-apoptosis proteins.…”

Section: Discussionmentioning

confidence: 99%

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Differential Impact of MouseRad9Deletion on Ionizing Radiation-Induced Bystander Effects

et al. 2005

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The cellular response to ionizing radiation is not limited to cells irradiated directly but can be demonstrated in neighboring "bystander" populations. The ability of mouse embryonic stem (ES) cells to express a bystander effect and the role of the radioresistance gene Rad9 were tested. Mouse ES cells differing in Rad9 status were exposed to broad-beam 125 keV/ μm 3 He α particles. All populations, when confluent, demonstrated a dose-independent bystander effect with respect to cell killing, and the Rad9 − / − genotype did not selectively alter that response or cell killing after direct exposure to this high-LET radiation. In contrast, relative to Rad9 + / + cells, the homozygous mutant was sensitive to direct exposure to α particles when in log phase, providing evidence of a role for Rad9 in repair of potentially lethal damage. Direct exposure to α particles induced an increase in the frequency of apoptosis and micronucleus formation, regardless of Rad9 status, although the null mutant showed high spontaneous levels of both end points. All populations demonstrated α-particle-induced bystander apoptosis, but that effect was most prominent in Rad9 − / − cells. Minimal α-particle induction of micro-nuclei in bystander cells was observed, except for the Rad9 − / − mutant, where a significant increase above background was detected. Therefore, the Rad9 null mutation selectively sensitizes mouse ES cells to spontaneous and high-LET radiation-induced bystander apoptosis and micronucleus formation, but it has much less impact on cell killing by direct or bystander α-particle exposure. Results are presented in the context of defining the function of Rad9 in the cellular response to radiation and its differential effects on individual bystander end points.

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Section: Discussionmentioning

confidence: 99%

“…The rad9 gene was first identified in the fission yeast Schizosaccharomyces pombe (8,9), and subsequently orthologs were found in humans [RAD9 (10)] and mice [Rad9 (11)]. The encoded protein plays a critical role in multiple pathways that respond to DNA damage.…”

Section: Introductionmentioning

confidence: 99%

Differential Impact of MouseRad9Deletion on Ionizing Radiation-Induced Bystander Effects

et al. 2005

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“…Mammalian versions of several have been isolated, providing strong evidence that checkpoint control mechanisms are also highly conserved (24,40,44,47,53,61,62,67,73). Human and mouse cDNAs encoding proteins highly homologous to the S. pombe rad9 gene product have been cloned (30,47). The cDNAs are capable of partially rescuing the hydroxyurea sensitivity, radi-osensitivity, and associated checkpoint deficiencies of S. pombe rad9 mutant cells.…”

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Deletion of Mouse Rad9 Causes Abnormal Cellular Responses to DNA Damage, Genomic Instability, and Embryonic Lethality

Hopkins

Auerbach

Wang

et al. 2004

Molecular and Cellular Biology

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The fission yeast Schizosaccharomyces pombe rad9 gene promotes cell survival through activation of cell cycle checkpoints induced by DNA damage. Mouse embryonic stem cells with a targeted deletion of Mrad9, the mouse ortholog of this gene, were created to evaluate its function in mammals. Mrad9؊/؊ cells demonstrated a marked increase in spontaneous chromosome aberrations and HPRT mutations, indicating a role in the maintenance of genomic integrity. These cells were also extremely sensitive to UV light, gamma rays, and hydroxyurea, and heterozygotes were somewhat sensitive to the last two agents relative to Mrad9 ؉/؉ controls. Mrad9 ؊/؊ cells could initiate but not maintain gamma-ray-induced G 2 delay and retained the ability to delay DNA synthesis rapidly after UV irradiation, suggesting that checkpoint abnormalities contribute little to the radiosensitivity observed. Ectopic expression of Mrad9 or human HRAD9 complemented Mrad9 ؊/؊ cell defects, indicating that the gene has radioresponse and genomic maintenance functions that are evolutionarily conserved. Mrad9 ؉/؊ mice were generated, but heterozygous intercrosses failed to yield Mrad9 ؊/؊ pups, since embryos died at midgestation. Furthermore, Mrad9 ؊/؊ mouse embryo fibroblasts were not viable. These investigations establish Mrad9 as a key mammalian genetic element of pathways that regulate the cellular response to DNA damage, maintenance of genomic integrity, and proper embryonic development.

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“…Consistent with the possibility that the mammalian 9-1-1 complex plays critical roles during gametogenesis, 9-1-1 subunits are highly expressed in the mouse testis, 14,15,33,[65][66][67] and both RAD1 and RAD9A localize to meiotic chromosomes. 15,33 Furthermore, Hus1 gene expression is reduced in the mouse testis in the absence of SPO11, 68 although it remains possible that this indicates that HUS1 function requires progression further into meiotic prophase I rather than a role specifically in meiotic recombination.…”

Section: Rad9a and Hus1 Are Critical For Mammalian Fertilitymentioning

confidence: 99%

Clamping down on mammalian meiosis

et al. 2013

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T he RAD9A-RAD1-HUS1 (9-1-1) complex is a PCNA-like heterotrimeric clamp that binds damaged DNA to promote cell cycle checkpoint signaling and DNA repair. While various 9-1-1 functions in mammalian somatic cells have been established, mounting evidence from lower eukaryotes predicts critical roles in meiotic germ cells as well. This was investigated in 2 recent studies in which the 9-1-1 complex was disrupted specifically in the mouse male germline through conditional deletion of Rad9a or Hus1. Loss of these clamp subunits led to severely impaired fertility and meiotic defects, including faulty DNA double-strand break repair. While 9-1-1 is critical for ATR kinase activation in somatic cells, these studies did not reveal major defects in ATR checkpoint pathway signaling in meiotic cells. Intriguingly, this new work identified separable roles for 9-1-1 subunits, namely RAD9A-and HUS1-independent roles for RAD1. Based on these studies and the high-level expression of the paralogous proteins RAD9B and HUS1B in testis, we propose a model in which multiple alternative 9-1-1 clamps function during mammalian meiosis to ensure genome maintenance in the germline.

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Molecular cloning and tissue-specific expression ofMrad9, a murine orthologue of theSchizosaccharomyces pombe rad9+ checkpoint control gene

Cited by 15 publications

References 31 publications

Differential Impact of MouseRad9Deletion on Ionizing Radiation-Induced Bystander Effects

Differential Impact of MouseRad9Deletion on Ionizing Radiation-Induced Bystander Effects

Deletion of Mouse Rad9 Causes Abnormal Cellular Responses to DNA Damage, Genomic Instability, and Embryonic Lethality

Clamping down on mammalian meiosis

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