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Novel inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 identified using a cell-based screen in yeast.
Cancer Res. 2001 May 15; 61(10):4175-83.CR

Abstract

Multicellular organisms must have means of preserving their genomic integrity or face catastrophic consequences such as uncontrolled cell proliferation or massive cell death. One response is a modification of nuclear proteins by the addition and removal of polymers of ADP-ribose that modulate the properties of DNA-binding proteins involved in DNA repair and metabolism. These ADP-ribose units are added by poly(ADP-ribose) polymerase (PARP) and removed by poly(ADP-ribose) glycohydrolase. Although budding yeast Saccharomyces cerevisiae does not possess proteins with significant sequence similarity to the human PARP family of proteins, we identified novel small molecule inhibitors against two family members, PARP1 and PARP2, using a cell-based assay in yeast. The assay was based on the reversal of growth inhibition caused by the heterologous expression of either PARP1 or PARP2. Validation of the assay was achieved by showing that the growth inhibition was relieved by a mutation in a single residue in the catalytic site of PARP1 or PARP2 or exposure of yeast to a known PARP1 inhibitor, 6(5H)-phenanthridinone. In separate experiments, when a putative protein regulator of PARP activity, human poly(ADP-ribose) glycohydrolase, was coexpressed with PARP1 or PARP2, yeast growth was restored. Finally, the inhibitors identified by screening the yeast assay are active in a mammalian PARP biochemical assay and inhibit PARP1 and PARP2 activity in yeast cell extracts. Thus, our data reflect the strength of using yeast to identify small molecule inhibitors of therapeutically relevant gene families, including those that are not found in yeast, such as PARP. The resultant inhibitors have two critical uses (a) as leads for drug development and (b) as tools to dissect cellular function.

Authors+Show Affiliations

Perkins E
Iconix Pharmaceuticals, 320 Logue Avenue, Mountain View, CA 94043, USA.
Sun D
No affiliation info available
Nguyen A
No affiliation info available
Tulac S
No affiliation info available
Francesco M
No affiliation info available
Tavana H
No affiliation info available
Nguyen H
No affiliation info available
Tugendreich S
No affiliation info available
Barthmaier P
No affiliation info available
Couto J
No affiliation info available
Yeh E
No affiliation info available
Thode S
No affiliation info available
Jarnagin K
No affiliation info available
Jain A
No affiliation info available
Morgans D
No affiliation info available
Melese T
No affiliation info available

MeSH

ATP-Binding Cassette TransportersBinding SitesDrug Evaluation, PreclinicalEnzyme InhibitorsGene ExpressionHumansIsoenzymesMembrane ProteinsMutationPhenanthrenesPoly(ADP-ribose) Polymerase InhibitorsPoly(ADP-ribose) PolymerasesSaccharomyces cerevisiaeSaccharomyces cerevisiae Proteins

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11358842

Citation

Perkins, E, et al. "Novel Inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 Identified Using a Cell-based Screen in Yeast." Cancer Research, vol. 61, no. 10, 2001, pp. 4175-83.
Perkins E, Sun D, Nguyen A, et al. Novel inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 identified using a cell-based screen in yeast. Cancer Res. 2001;61(10):4175-83.
Perkins, E., Sun, D., Nguyen, A., Tulac, S., Francesco, M., Tavana, H., Nguyen, H., Tugendreich, S., Barthmaier, P., Couto, J., Yeh, E., Thode, S., Jarnagin, K., Jain, A., Morgans, D., & Melese, T. (2001). Novel inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 identified using a cell-based screen in yeast. Cancer Research, 61(10), 4175-83.
Perkins E, et al. Novel Inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 Identified Using a Cell-based Screen in Yeast. Cancer Res. 2001 May 15;61(10):4175-83. PubMed PMID: 11358842.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Novel inhibitors of poly(ADP-ribose) polymerase/PARP1 and PARP2 identified using a cell-based screen in yeast. AU - Perkins,E, AU - Sun,D, AU - Nguyen,A, AU - Tulac,S, AU - Francesco,M, AU - Tavana,H, AU - Nguyen,H, AU - Tugendreich,S, AU - Barthmaier,P, AU - Couto,J, AU - Yeh,E, AU - Thode,S, AU - Jarnagin,K, AU - Jain,A, AU - Morgans,D, AU - Melese,T, PY - 2001/5/19/pubmed PY - 2001/6/23/medline PY - 2001/5/19/entrez SP - 4175 EP - 83 JF - Cancer research JO - Cancer Res VL - 61 IS - 10 N2 - Multicellular organisms must have means of preserving their genomic integrity or face catastrophic consequences such as uncontrolled cell proliferation or massive cell death. One response is a modification of nuclear proteins by the addition and removal of polymers of ADP-ribose that modulate the properties of DNA-binding proteins involved in DNA repair and metabolism. These ADP-ribose units are added by poly(ADP-ribose) polymerase (PARP) and removed by poly(ADP-ribose) glycohydrolase. Although budding yeast Saccharomyces cerevisiae does not possess proteins with significant sequence similarity to the human PARP family of proteins, we identified novel small molecule inhibitors against two family members, PARP1 and PARP2, using a cell-based assay in yeast. The assay was based on the reversal of growth inhibition caused by the heterologous expression of either PARP1 or PARP2. Validation of the assay was achieved by showing that the growth inhibition was relieved by a mutation in a single residue in the catalytic site of PARP1 or PARP2 or exposure of yeast to a known PARP1 inhibitor, 6(5H)-phenanthridinone. In separate experiments, when a putative protein regulator of PARP activity, human poly(ADP-ribose) glycohydrolase, was coexpressed with PARP1 or PARP2, yeast growth was restored. Finally, the inhibitors identified by screening the yeast assay are active in a mammalian PARP biochemical assay and inhibit PARP1 and PARP2 activity in yeast cell extracts. Thus, our data reflect the strength of using yeast to identify small molecule inhibitors of therapeutically relevant gene families, including those that are not found in yeast, such as PARP. The resultant inhibitors have two critical uses (a) as leads for drug development and (b) as tools to dissect cellular function. SN - 0008-5472 UR - https://www.unboundmedicine.com/medline/citation/11358842/Novel_inhibitors_of_poly_ADP_ribose__polymerase/PARP1_and_PARP2_identified_using_a_cell_based_screen_in_yeast_ DB - PRIME DP - Unbound Medicine ER -