SUPLEMENTARY MATERIAL BIOMETALS Role and regulation of ferritin-like proteins in iron homeostasis and oxidative stress survival of Caulobacter crescentus.

Apresentação em tema: "SUPLEMENTARY MATERIAL BIOMETALS Role and regulation of ferritin-like proteins in iron homeostasis and oxidative stress survival of Caulobacter crescentus."— Transcrição da apresentação:

1 SUPLEMENTARY MATERIAL BIOMETALS Role and regulation of ferritin-like proteins in iron homeostasis and oxidative stress survival of Caulobacter crescentus Ivan Gonçalves de Castro Ferreira a,1, Mirian Molnar Rodrigues a,1, José Freire da Silva Neto b, Ricardo Ruiz Mazzon c & Marilis do Valle Marques a * a Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1374, 05508-900 São Paulo, SP, Brazil b Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo - Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto, SP, Brazil c Departamento de Microbiologia, Imunologia e Parasitologia, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina – Campus Universitário da Trindade, 88040-900, Florianópolis, SC, Brazil 1 These authors contributed equally to this work *Corresponding author. Mailing address: Av. Prof. Lineu Prestes, 1374, 05508-900 São Paulo, SP, Brazil. Phone: 55-11-3091-7299; FAX: 55-11-3091-7354. E-mail: mvmarque@usp.brmvmarque@usp.br

2  dps 1.7 kb 1650 bp 2000 bp wt dps 1650 bp 2000 bp  bfr 1.5 kb wt bfr dps dps/ bfr 1.5 kb 1650 bp 2000 bp  dps/  bfr Figure S1: Confirmation of the deleted strains. Total genomic DNA was used as template in PCR reactions with primers dps1/dps4 (for dps deletion), or bfr1/bfr4 (for bfr deletion in NA1000 and dps backgrounds). For the single mutants genomic DNA from the wild type strain NA1000 was used, and for the double mutant DNA from the dps mutant was used. The sizes of the bands corresponding to the amplicon of the deleted gene are indicated. The DNA markers were run on the same gels as the amplified bands.

3 Figure S2: Expression of dps/lacZ fusion. Plasmid pMMR02 carrying a transcriptional fusion of dps promoter region to the lacZ reporter gene was introduced into NA1000 (wt) and SP3811 (  oxyR) strains. Cultures of these strains were diluted (OD600nm = 0.1), and aliquots were taken for  -galactosidase activity assays at mid-log phase before (time zero) and at several time points after addition of 60  M of H 2 O 2 (as indicated). One asterisk indicates statistical difference between different conditions for a given strain (*) or between different strains at the same condition (**) according to parametric unpaired t-test (p<0.05). wt  oxyR Time (min) 0 5 15 300 5 15 30 * ** * *