Identification of secondary targets of N-containing bisphosphonates in mammalian cells via parallel competition analysis of the barcoded yeast deletion collectionReportar como inadecuado




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Genome Biology

, 10:R93

First Online: 10 September 2009Received: 14 May 2009Revised: 16 July 2009Accepted: 10 September 2009

Abstract

BackgroundNitrogen-containing bisphosphonates are the elected drugs for the treatment of diseases in which excessive bone resorption occurs, for example, osteoporosis and cancer-induced bone diseases. The only known target of nitrogen-containing bisphosphonates is farnesyl pyrophosphate synthase, which ensures prenylation of prosurvival proteins, such as Ras. However, it is likely that the action of nitrogen-containing bisphosphonates involves additional unknown mechanisms. To identify novel targets of nitrogen-containing bisphosphonates, we used a genome-wide high-throughput screening in which 5,936 Saccharomyces cerevisiae heterozygote barcoded mutants were grown competitively in the presence of sub-lethal doses of three nitrogen-containing bisphosphonates risedronate, alendronate and ibandronate. Strains carrying deletions in genes encoding potential drug targets show a variation of the intensity of their corresponding barcodes on the hybridization array over the time.

ResultsWith this approach, we identified novel targets of nitrogen-containing bisphosphonates, such as tubulin cofactor B and ASK-DBF4 Activator of S-phase kinase. The up-regulation of tubulin cofactor B may explain some previously unknown effects of nitrogen-containing bisphosphonates on microtubule dynamics and organization. As nitrogen-containing bisphosphonates induce extensive DNA damage, we also document the role of DBF4 as a key player in nitrogen-containing bisphosphonate-induced cytotoxicity, thus explaining the effects on the cell-cycle.

ConclusionsThe dataset obtained from the yeast screen was validated in a mammalian system, allowing the discovery of new biological processes involved in the cellular response to nitrogen-containing bisphosphonates and opening up opportunities for development of new anticancer drugs.

AbbreviationsALEalendronate

DMEMDulbecco-s modified Eagle-s medium

FPPSfarnesyl pyrophosphate synthase

GGPPgeranylgeranyl pyrophosphate

IBAibandronate

N-BPsnitrogen bisphosphonate

ORFopen reading frame

QDquick disappearing

RISrisedronate

siRNAsmall interfering RNA

TBCBtubulin cofactor B.

Electronic supplementary materialThe online version of this article doi:10.1186-gb-2009-10-9-r93 contains supplementary material, which is available to authorized users.

Gianluca Tell and Daniela Delneri contributed equally to this work.

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Autor: Nicoletta Bivi - Milena Romanello - Richard Harrison - Ian Clarke - David C Hoyle - Luigi Moro - Fulvia Ortolani - Antonel

Fuente: https://link.springer.com/







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