000074972 001__ 74972
000074972 005__ 20200117221609.0
000074972 0247_ $$2doi$$a10.3389/fmicb.2018.01659
000074972 0248_ $$2sideral$$a107619
000074972 037__ $$aART-2018-107619
000074972 041__ $$aeng
000074972 100__ $$aGarcía, M.T.
000074972 245__ $$aBoldine-derived Alkaloids inhibit the activity of DNA topoisomerase I and growth of Mycobacterium tuberculosis
000074972 260__ $$c2018
000074972 5060_ $$aAccess copy available to the general public$$fUnrestricted
000074972 5203_ $$aThe spread of multidrug-resistant isolates of Mycobacterium tuberculosis requires the discovery of new drugs directed to new targets. In this study, we investigated the activity of two boldine-derived alkaloids, seconeolitsine (SCN) and N-methyl-seconeolitsine (N-SCN), against M. tuberculosis. These compounds have been shown to target DNA topoisomerase I enzyme and inhibit growth of Streptococcus pneumoniae. Both SCN and N-SCN inhibited M. tuberculosis growth at 1.95-15.6 µM, depending on the strain. In M. smegmatis this inhibitory effect correlated with the amount of topoisomerase I in the cell, hence demonstrating that this enzyme is the target for these alkaloids in mycobacteria. The gene coding for topoisomerase I of strain H37Rv (MtbTopoI) was cloned into pQE1 plasmid of Escherichia coli. MtbTopoI was overexpressed with an N-terminal 6-His-tag and purified by affinity chromatography. In vitro inhibition of MtbTopoI activity by SCN and N-SCN was tested using a plasmid relaxation assay. Both SCN and N-SCN inhibited 50% of the enzymatic activity at 5.6 and 8.4 µM, respectively. Cleavage of single-stranded DNA was also inhibited with SCN. The effects on DNA supercoiling were also evaluated in vivo in plasmid-containing cultures of M. tuberculosis. Plasmid supercoiling densities were -0.060 in cells untreated or treated with boldine, and -0.072 in 1 × MIC N-SCN treated cells, respectively, indicating that the plasmid became hypernegatively supercoiled in the presence of N-SCN. Altogether, these results demonstrate that the M. tuberculosis topoisomerase I enzyme is an attractive drug target, and that SCN and N-SCN are promising lead compounds for drug development.
000074972 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/BIO2017-82951-R$$9info:eu-repo/grantAgreement/ES/MINECO/BIO2009-09405$$9info:eu-repo/grantAgreement/EC/FP7/260872/EU/More Medicines for Tuberculosis/MM4TB
000074972 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000074972 590__ $$a4.259$$b2018
000074972 591__ $$aMICROBIOLOGY$$b32 / 133 = 0.241$$c2018$$dQ1$$eT1
000074972 592__ $$a1.633$$b2018
000074972 593__ $$aMicrobiology (medical)$$c2018$$dQ1
000074972 593__ $$aMicrobiology$$c2018$$dQ1
000074972 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000074972 700__ $$aCarreño, D.
000074972 700__ $$aTirado-Vélez, J.M.
000074972 700__ $$aFerrándiz, M.J.
000074972 700__ $$aRodrigues, L.
000074972 700__ $$aGracia, B.$$uUniversidad de Zaragoza
000074972 700__ $$aAmblar, M.
000074972 700__ $$0(orcid)0000-0003-2076-844X$$aAinsa, J.A.$$uUniversidad de Zaragoza
000074972 700__ $$ade la Campa, A.G.
000074972 7102_ $$11008$$2630$$aUniversidad de Zaragoza$$bDpto. Microb.Med.Pr.,Sal.Públ.$$cÁrea Microbiología
000074972 7102_ $$11008$$2X$$aUniversidad de Zaragoza$$bDpto. Microb.Med.Pr.,Sal.Públ.$$cProy. investigación HQA
000074972 773__ $$g9, JUL (2018), 1659 [9 pp]$$pFront. microbiol.$$tFRONTIERS IN MICROBIOLOGY$$x1664-302X
000074972 8564_ $$s748389$$uhttps://zaguan.unizar.es/record/74972/files/texto_completo.pdf$$yVersión publicada
000074972 8564_ $$s28488$$uhttps://zaguan.unizar.es/record/74972/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000074972 909CO $$ooai:zaguan.unizar.es:74972$$particulos$$pdriver
000074972 951__ $$a2020-01-17-21:47:59
000074972 980__ $$aARTICLE