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Caenorhabditis elegans protein arginine methyltransferase PRMT-5 negatively regulates DNA damage-induced apoptosis.

Yang M, Sun J, Sun X, Shen Q, Gao Z, Yang C - PLoS Genet. (2009)

Bottom Line: PRMT-5 forms a complex with both CEP-1 and CBP-1 and can methylate the latter.Importantly, down-regulation of cbp-1 significantly suppresses DNA damage-induced egl-1 expression and apoptosis in prmt-5 mutant worms.These findings suggest that PRMT-5 likely represses CEP-1 transcriptional activity through CBP-1, which represents a novel regulatory mechanism of p53-dependent apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
Arginine methylation of histone and non-histone proteins is involved in transcription regulation and many other cellular processes. Nevertheless, whether such protein modification plays a regulatory role during apoptosis remains largely unknown. Here we report that the Caenorhabditis elegans homolog of mammalian type II arginine methyltransferase PRMT5 negatively regulates DNA damage-induced apoptosis. We show that inactivation of C. elegans prmt-5 leads to excessive apoptosis in germline following ionizing irradiation, which is due to a CEP-1/p53-dependent up-regulation of the cell death initiator EGL-1. Moreover, we provide evidence that CBP-1, the worm ortholog of human p300/CBP, functions as a cofactor of CEP-1. PRMT-5 forms a complex with both CEP-1 and CBP-1 and can methylate the latter. Importantly, down-regulation of cbp-1 significantly suppresses DNA damage-induced egl-1 expression and apoptosis in prmt-5 mutant worms. These findings suggest that PRMT-5 likely represses CEP-1 transcriptional activity through CBP-1, which represents a novel regulatory mechanism of p53-dependent apoptosis.

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Epitasis analysis of prmt-5-mediated apoptosis.(A) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), ced-3(n717), egl-1(n1084 n3082) single mutants and prmt-5(gk357);ced-3(n717) as well as prmt-5(gk357);egl-1(n1084 n3082) double mutants. Young adult animals were irradiated with γ-ray of 120 Gy and analyzed 36 h post irradiation. Germ cell corpses from one gonad arm of each animal were scored for at least 20 animals. Error bars represent SEM. (B) Quantification of germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, ced-4(n1162) and ced-9(n1950gf) mutants following γ-irradiation. Worms were irradiated and germ cell corpses were scored and analyzed as in (A). (C) Quantification of IR-induced germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, hus-1(op241), mrt-2(e2663) and clk-2(mn159) animals. Worm treatment and germ cell corpse analysis were performed as in (A). (D) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), hus-1(op241) single mutants and hus-1(op241);prmt-5(gk357) double mutants. Worm treatment and germ cell corpse analysis were performed as in (A).
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pgen-1000514-g002: Epitasis analysis of prmt-5-mediated apoptosis.(A) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), ced-3(n717), egl-1(n1084 n3082) single mutants and prmt-5(gk357);ced-3(n717) as well as prmt-5(gk357);egl-1(n1084 n3082) double mutants. Young adult animals were irradiated with γ-ray of 120 Gy and analyzed 36 h post irradiation. Germ cell corpses from one gonad arm of each animal were scored for at least 20 animals. Error bars represent SEM. (B) Quantification of germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, ced-4(n1162) and ced-9(n1950gf) mutants following γ-irradiation. Worms were irradiated and germ cell corpses were scored and analyzed as in (A). (C) Quantification of IR-induced germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, hus-1(op241), mrt-2(e2663) and clk-2(mn159) animals. Worm treatment and germ cell corpse analysis were performed as in (A). (D) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), hus-1(op241) single mutants and hus-1(op241);prmt-5(gk357) double mutants. Worm treatment and germ cell corpse analysis were performed as in (A).

Mentions: Several lines of evidence have shown that germ cell apoptosis induced by DNA damage requires the core cell death pathway because mutations of genes essential for programmed cell death, including ced-3, ced-4, ced-9 and egl-1, block such cell death. To determine whether the strong increase of IR-induced germ cell apoptosis in prmt-5(gk357) mutants is dependent on the core cell death pathway, we generated prmt-5(gk357);ced-3(n717) and prmt-5(gk357);egl-1(n1084 n3082) double mutants and found that germ cell apoptosis was barely induced by IR in these worms (Figure 2A). Moreover, in prmt-5 RNAi-treated ced-4(n1162) loss-of-function and ced-9(n1950) gain-of-function mutants, IR-induced germ cell apoptosis was either abrogated or strongly suppressed as compared with that in prmt-5 RNAi-treated wild-type animals (Figure 2B). These results suggest that prmt-5 acts through the core cell death pathway to regulate DNA damage-induced apoptosis.


Caenorhabditis elegans protein arginine methyltransferase PRMT-5 negatively regulates DNA damage-induced apoptosis.

Yang M, Sun J, Sun X, Shen Q, Gao Z, Yang C - PLoS Genet. (2009)

Epitasis analysis of prmt-5-mediated apoptosis.(A) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), ced-3(n717), egl-1(n1084 n3082) single mutants and prmt-5(gk357);ced-3(n717) as well as prmt-5(gk357);egl-1(n1084 n3082) double mutants. Young adult animals were irradiated with γ-ray of 120 Gy and analyzed 36 h post irradiation. Germ cell corpses from one gonad arm of each animal were scored for at least 20 animals. Error bars represent SEM. (B) Quantification of germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, ced-4(n1162) and ced-9(n1950gf) mutants following γ-irradiation. Worms were irradiated and germ cell corpses were scored and analyzed as in (A). (C) Quantification of IR-induced germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, hus-1(op241), mrt-2(e2663) and clk-2(mn159) animals. Worm treatment and germ cell corpse analysis were performed as in (A). (D) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), hus-1(op241) single mutants and hus-1(op241);prmt-5(gk357) double mutants. Worm treatment and germ cell corpse analysis were performed as in (A).
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pgen-1000514-g002: Epitasis analysis of prmt-5-mediated apoptosis.(A) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), ced-3(n717), egl-1(n1084 n3082) single mutants and prmt-5(gk357);ced-3(n717) as well as prmt-5(gk357);egl-1(n1084 n3082) double mutants. Young adult animals were irradiated with γ-ray of 120 Gy and analyzed 36 h post irradiation. Germ cell corpses from one gonad arm of each animal were scored for at least 20 animals. Error bars represent SEM. (B) Quantification of germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, ced-4(n1162) and ced-9(n1950gf) mutants following γ-irradiation. Worms were irradiated and germ cell corpses were scored and analyzed as in (A). (C) Quantification of IR-induced germ cell apoptosis in control RNAi- and prmt-5 RNAi-treated N2, hus-1(op241), mrt-2(e2663) and clk-2(mn159) animals. Worm treatment and germ cell corpse analysis were performed as in (A). (D) Quantification of IR-induced germ cell apoptosis in N2, prmt-5(gk357), hus-1(op241) single mutants and hus-1(op241);prmt-5(gk357) double mutants. Worm treatment and germ cell corpse analysis were performed as in (A).
Mentions: Several lines of evidence have shown that germ cell apoptosis induced by DNA damage requires the core cell death pathway because mutations of genes essential for programmed cell death, including ced-3, ced-4, ced-9 and egl-1, block such cell death. To determine whether the strong increase of IR-induced germ cell apoptosis in prmt-5(gk357) mutants is dependent on the core cell death pathway, we generated prmt-5(gk357);ced-3(n717) and prmt-5(gk357);egl-1(n1084 n3082) double mutants and found that germ cell apoptosis was barely induced by IR in these worms (Figure 2A). Moreover, in prmt-5 RNAi-treated ced-4(n1162) loss-of-function and ced-9(n1950) gain-of-function mutants, IR-induced germ cell apoptosis was either abrogated or strongly suppressed as compared with that in prmt-5 RNAi-treated wild-type animals (Figure 2B). These results suggest that prmt-5 acts through the core cell death pathway to regulate DNA damage-induced apoptosis.

Bottom Line: PRMT-5 forms a complex with both CEP-1 and CBP-1 and can methylate the latter.Importantly, down-regulation of cbp-1 significantly suppresses DNA damage-induced egl-1 expression and apoptosis in prmt-5 mutant worms.These findings suggest that PRMT-5 likely represses CEP-1 transcriptional activity through CBP-1, which represents a novel regulatory mechanism of p53-dependent apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
Arginine methylation of histone and non-histone proteins is involved in transcription regulation and many other cellular processes. Nevertheless, whether such protein modification plays a regulatory role during apoptosis remains largely unknown. Here we report that the Caenorhabditis elegans homolog of mammalian type II arginine methyltransferase PRMT5 negatively regulates DNA damage-induced apoptosis. We show that inactivation of C. elegans prmt-5 leads to excessive apoptosis in germline following ionizing irradiation, which is due to a CEP-1/p53-dependent up-regulation of the cell death initiator EGL-1. Moreover, we provide evidence that CBP-1, the worm ortholog of human p300/CBP, functions as a cofactor of CEP-1. PRMT-5 forms a complex with both CEP-1 and CBP-1 and can methylate the latter. Importantly, down-regulation of cbp-1 significantly suppresses DNA damage-induced egl-1 expression and apoptosis in prmt-5 mutant worms. These findings suggest that PRMT-5 likely represses CEP-1 transcriptional activity through CBP-1, which represents a novel regulatory mechanism of p53-dependent apoptosis.

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