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Double-strand break repair deficiency in NONO knockout murine embryonic fibroblasts and compensation by spontaneous upregulation of the PSPC1 paralog.

Li S, Li Z, Shu FJ, Xiong H, Phillips AC, Dynan WS - Nucleic Acids Res. (2014)

Bottom Line: We tested whether NONO and PSPC1 might also affect repair indirectly by influencing mRNA levels for other DSB repair genes.Of 12 genes tested, none were downregulated, and several were upregulated.Thus, NONO or related proteins are critical for DSB repair, NONO and PSPC1 are functional homologs with partially interchangeable functions and a compensatory response involving PSPC1 blunts the effect of NONO deficiency.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Emory University, Atlanta, GA 30322, USA Department of Biochemistry, Emory University, Atlanta, GA 30322, USA Institute of Molecular Medicine and Genetics, Georgia Regents University, Augusta, GA 30912, USA.

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Relative mRNA expression for other DSB repair genes. Vectors encoding control or PSPC1 miRNA were introduced by electroporation into wild-type (+/0) or NONO-deficient (gt/0) MEFs. Expression was measured using Taqman probes with Gapdh as an internal reference. Data are normalized to expression in control miRNA-transfected wild-type MEFs. Values are mean of three independent biological replicates with standard deviation shown. Genes are grouped according to the presence of a DBHS motif, participation in canonical NHEJ (C-NHEJ), participation in alternative NHEJ (A-NHEJ) or participation in homology-directed repair (HR), respectively (**P < 0.01 and *P < 0.05).
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Figure 6: Relative mRNA expression for other DSB repair genes. Vectors encoding control or PSPC1 miRNA were introduced by electroporation into wild-type (+/0) or NONO-deficient (gt/0) MEFs. Expression was measured using Taqman probes with Gapdh as an internal reference. Data are normalized to expression in control miRNA-transfected wild-type MEFs. Values are mean of three independent biological replicates with standard deviation shown. Genes are grouped according to the presence of a DBHS motif, participation in canonical NHEJ (C-NHEJ), participation in alternative NHEJ (A-NHEJ) or participation in homology-directed repair (HR), respectively (**P < 0.01 and *P < 0.05).

Mentions: Results, shown in Figure 6, indicate that the single greatest effect of NONO deficiency was upregulation of PSPC1, consistent with observations at the protein level. The upregulation of PSPC1 was partially reversed by transfection with PSPC1 miRNA, although suppression is incomplete, likely because the population consists of a mixture of transfected and non-transfected cells. There was also a smaller, but significant, upregulation of SFPQ mRNA, although these had not been seen at the protein level in Figure 3.


Double-strand break repair deficiency in NONO knockout murine embryonic fibroblasts and compensation by spontaneous upregulation of the PSPC1 paralog.

Li S, Li Z, Shu FJ, Xiong H, Phillips AC, Dynan WS - Nucleic Acids Res. (2014)

Relative mRNA expression for other DSB repair genes. Vectors encoding control or PSPC1 miRNA were introduced by electroporation into wild-type (+/0) or NONO-deficient (gt/0) MEFs. Expression was measured using Taqman probes with Gapdh as an internal reference. Data are normalized to expression in control miRNA-transfected wild-type MEFs. Values are mean of three independent biological replicates with standard deviation shown. Genes are grouped according to the presence of a DBHS motif, participation in canonical NHEJ (C-NHEJ), participation in alternative NHEJ (A-NHEJ) or participation in homology-directed repair (HR), respectively (**P < 0.01 and *P < 0.05).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150768&req=5

Figure 6: Relative mRNA expression for other DSB repair genes. Vectors encoding control or PSPC1 miRNA were introduced by electroporation into wild-type (+/0) or NONO-deficient (gt/0) MEFs. Expression was measured using Taqman probes with Gapdh as an internal reference. Data are normalized to expression in control miRNA-transfected wild-type MEFs. Values are mean of three independent biological replicates with standard deviation shown. Genes are grouped according to the presence of a DBHS motif, participation in canonical NHEJ (C-NHEJ), participation in alternative NHEJ (A-NHEJ) or participation in homology-directed repair (HR), respectively (**P < 0.01 and *P < 0.05).
Mentions: Results, shown in Figure 6, indicate that the single greatest effect of NONO deficiency was upregulation of PSPC1, consistent with observations at the protein level. The upregulation of PSPC1 was partially reversed by transfection with PSPC1 miRNA, although suppression is incomplete, likely because the population consists of a mixture of transfected and non-transfected cells. There was also a smaller, but significant, upregulation of SFPQ mRNA, although these had not been seen at the protein level in Figure 3.

Bottom Line: We tested whether NONO and PSPC1 might also affect repair indirectly by influencing mRNA levels for other DSB repair genes.Of 12 genes tested, none were downregulated, and several were upregulated.Thus, NONO or related proteins are critical for DSB repair, NONO and PSPC1 are functional homologs with partially interchangeable functions and a compensatory response involving PSPC1 blunts the effect of NONO deficiency.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Emory University, Atlanta, GA 30322, USA Department of Biochemistry, Emory University, Atlanta, GA 30322, USA Institute of Molecular Medicine and Genetics, Georgia Regents University, Augusta, GA 30912, USA.

Show MeSH
Related in: MedlinePlus