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TRIM3, a tumor suppressor linked to regulation of p21(Waf1/Cip1.).

Liu Y, Raheja R, Yeh N, Ciznadija D, Pedraza AM, Ozawa T, Hukkelhoven E, Erdjument-Bromage H, Tempst P, Gauthier NP, Brennan C, Holland EC, Koff A - Oncogene (2013)

Bottom Line: The TRIM family of genes is largely studied because of their roles in development, differentiation and host cell antiviral defenses; however, roles in cancer biology are emerging.Furthermore, TRIM3 can bind to the cdk inhibitor p21(WAF1/CIP1).Thus, we conclude that TRIM3 is a tumor suppressor mapping to chromosome 11p15.5 and that it might block tumor growth by sequestering p21 and preventing it from facilitating the accumulation of cyclin D1-cdk4.

View Article: PubMed Central - PubMed

Affiliation: Programs in Molecular Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.

ABSTRACT
The TRIM family of genes is largely studied because of their roles in development, differentiation and host cell antiviral defenses; however, roles in cancer biology are emerging. Loss of heterozygosity of the TRIM3 locus in ∼20% of human glioblastomas raised the possibility that this NHL-domain containing member of the TRIM gene family might be a mammalian tumor suppressor. Consistent with this, reducing TRIM3 expression increased the incidence of and accelerated the development of platelet-derived growth factor -induced glioma in mice. Furthermore, TRIM3 can bind to the cdk inhibitor p21(WAF1/CIP1). Thus, we conclude that TRIM3 is a tumor suppressor mapping to chromosome 11p15.5 and that it might block tumor growth by sequestering p21 and preventing it from facilitating the accumulation of cyclin D1-cdk4.

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The association of TRIM3 with p21 is reduced in the presence of cyclin-cdk complexes binding to p21(A) Immunoblot. GST-p21 and GST-p21cy(-) were produced and purified from E. coli and the amount indicated above each lane (ng) was incubated with 0.5mg total cell extract obtained from myc-TRIM3 transfected 293T cells. myc-TRIM3 was subsequently immunoprecipitated and the amount of TRIM3 and p21 determined by immunoblotting as indicated to the right of each panel. (B) Extracts from myc-TRIM3 expressing 293T cells were passed through either p13-sepharose to deplete cyclin-cdk comlexes or sepharose CL-4B (mock). On the left, the amount of cyclin A, cyclin D1, cdk2 and cdk4 was measured by immunoblot to assess whether the depletion was successful. On the right, 12.5ng of recombinant substrate indicated above each lane was incubated with 0.25mg of the depleted extracts and recovered as described.
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Figure 8: The association of TRIM3 with p21 is reduced in the presence of cyclin-cdk complexes binding to p21(A) Immunoblot. GST-p21 and GST-p21cy(-) were produced and purified from E. coli and the amount indicated above each lane (ng) was incubated with 0.5mg total cell extract obtained from myc-TRIM3 transfected 293T cells. myc-TRIM3 was subsequently immunoprecipitated and the amount of TRIM3 and p21 determined by immunoblotting as indicated to the right of each panel. (B) Extracts from myc-TRIM3 expressing 293T cells were passed through either p13-sepharose to deplete cyclin-cdk comlexes or sepharose CL-4B (mock). On the left, the amount of cyclin A, cyclin D1, cdk2 and cdk4 was measured by immunoblot to assess whether the depletion was successful. On the right, 12.5ng of recombinant substrate indicated above each lane was incubated with 0.25mg of the depleted extracts and recovered as described.

Mentions: Given that p21 is a cyclin D-cdk4 assembly factor in the RCAS-PDGF-HA/nestin-TvA model [30], and TRIM3 can bind to p21 and suppress cell proliferation, we wanted to know if cyclin-cdks affected the interaction of TRIM3 and p21. We found that a mutant of p21 deficient in binding cyclin-cdk complexes bound better to TRIM3 than wild type p21 in extracts containing cyclin-cdk complexes (Fig. 8A). However the wild type p21 and the non-cyclin-cdk binding mutant bound equivalently if cyclin-cdk complexes were depleted from the extract (Fig. 8B). Thus, TRIM3 binds better to p21 that was not physically associated with cyclin-cdk complexes, and TRIM3 and cyclin-cdk complexes might compete with each other for p21.


TRIM3, a tumor suppressor linked to regulation of p21(Waf1/Cip1.).

Liu Y, Raheja R, Yeh N, Ciznadija D, Pedraza AM, Ozawa T, Hukkelhoven E, Erdjument-Bromage H, Tempst P, Gauthier NP, Brennan C, Holland EC, Koff A - Oncogene (2013)

The association of TRIM3 with p21 is reduced in the presence of cyclin-cdk complexes binding to p21(A) Immunoblot. GST-p21 and GST-p21cy(-) were produced and purified from E. coli and the amount indicated above each lane (ng) was incubated with 0.5mg total cell extract obtained from myc-TRIM3 transfected 293T cells. myc-TRIM3 was subsequently immunoprecipitated and the amount of TRIM3 and p21 determined by immunoblotting as indicated to the right of each panel. (B) Extracts from myc-TRIM3 expressing 293T cells were passed through either p13-sepharose to deplete cyclin-cdk comlexes or sepharose CL-4B (mock). On the left, the amount of cyclin A, cyclin D1, cdk2 and cdk4 was measured by immunoblot to assess whether the depletion was successful. On the right, 12.5ng of recombinant substrate indicated above each lane was incubated with 0.25mg of the depleted extracts and recovered as described.
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Related In: Results  -  Collection

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Show All Figures
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Figure 8: The association of TRIM3 with p21 is reduced in the presence of cyclin-cdk complexes binding to p21(A) Immunoblot. GST-p21 and GST-p21cy(-) were produced and purified from E. coli and the amount indicated above each lane (ng) was incubated with 0.5mg total cell extract obtained from myc-TRIM3 transfected 293T cells. myc-TRIM3 was subsequently immunoprecipitated and the amount of TRIM3 and p21 determined by immunoblotting as indicated to the right of each panel. (B) Extracts from myc-TRIM3 expressing 293T cells were passed through either p13-sepharose to deplete cyclin-cdk comlexes or sepharose CL-4B (mock). On the left, the amount of cyclin A, cyclin D1, cdk2 and cdk4 was measured by immunoblot to assess whether the depletion was successful. On the right, 12.5ng of recombinant substrate indicated above each lane was incubated with 0.25mg of the depleted extracts and recovered as described.
Mentions: Given that p21 is a cyclin D-cdk4 assembly factor in the RCAS-PDGF-HA/nestin-TvA model [30], and TRIM3 can bind to p21 and suppress cell proliferation, we wanted to know if cyclin-cdks affected the interaction of TRIM3 and p21. We found that a mutant of p21 deficient in binding cyclin-cdk complexes bound better to TRIM3 than wild type p21 in extracts containing cyclin-cdk complexes (Fig. 8A). However the wild type p21 and the non-cyclin-cdk binding mutant bound equivalently if cyclin-cdk complexes were depleted from the extract (Fig. 8B). Thus, TRIM3 binds better to p21 that was not physically associated with cyclin-cdk complexes, and TRIM3 and cyclin-cdk complexes might compete with each other for p21.

Bottom Line: The TRIM family of genes is largely studied because of their roles in development, differentiation and host cell antiviral defenses; however, roles in cancer biology are emerging.Furthermore, TRIM3 can bind to the cdk inhibitor p21(WAF1/CIP1).Thus, we conclude that TRIM3 is a tumor suppressor mapping to chromosome 11p15.5 and that it might block tumor growth by sequestering p21 and preventing it from facilitating the accumulation of cyclin D1-cdk4.

View Article: PubMed Central - PubMed

Affiliation: Programs in Molecular Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.

ABSTRACT
The TRIM family of genes is largely studied because of their roles in development, differentiation and host cell antiviral defenses; however, roles in cancer biology are emerging. Loss of heterozygosity of the TRIM3 locus in ∼20% of human glioblastomas raised the possibility that this NHL-domain containing member of the TRIM gene family might be a mammalian tumor suppressor. Consistent with this, reducing TRIM3 expression increased the incidence of and accelerated the development of platelet-derived growth factor -induced glioma in mice. Furthermore, TRIM3 can bind to the cdk inhibitor p21(WAF1/CIP1). Thus, we conclude that TRIM3 is a tumor suppressor mapping to chromosome 11p15.5 and that it might block tumor growth by sequestering p21 and preventing it from facilitating the accumulation of cyclin D1-cdk4.

Show MeSH
Related in: MedlinePlus