title: The BCG Moreau RD16 deletion inactivates a repressor reshaping transcription of an adjacent gene.
abstract: The Brazilian anti-tuberculosis vaccine strain Mycobacterium bovis bacillus Calmette-Guérin (BCG) BCG Moreau is unique in having a deletion of 7608 bp (RD16) that results in the truncation of a putative TetR transcriptional regulator, the ortholog of Mycobacterium tuberculosis rv3405c, BCG_M3439c. We investigated the effect of this truncation on the expression of the rv3406 ortholog (BCG_M3440), lying 81 bp downstream in the opposite orientation. RT-PCR and western blot experiments show that rv3406 mRNA and Rv3406 accumulate in BCG Moreau but not in BCG Pasteur (strain that bears an intact rv3405c), suggesting this to be a result of rv3405c truncation. Recombinant Rv3405c forms a complex with the rv3405c-rv3406 intergenic region, which contains a characteristic transcription factor binding site, showing it to have DNA binding activity. Complementation of M. bovis BCG Moreau with an intact copy of rv3405c abolishes Rv3406 accumulation. These results show that Rv3405c is a DNA binding protein that acts as a transcriptional repressor of rv3406.
authors: Laboratório de Bacteriologia, Centro de Referência Prof. Hélio Fraga, Escola Nacional de Saúde Pública Sergio Arouca, Fiocruz. Estr. de Curicica 2000, Jacarepaguá, 22780-194 Rio de Janeiro, RJ, Brazil. Electronic address: tecagalvao1@gmail.com.
authors: Teca Calcagno
authors: TC
authors: Galvão
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: crika@ioc.fiocruz.br.
authors: Cristiane Rodrigues
authors: CR
authors: Lima
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: leonardoh@ioc.fiocruz.br.
authors: Leonardo Henrique Ferreira
authors: LH
authors: Gomes
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: talitadp@ioc.fiocruz.br.
authors: Talita Duarte
authors: TD
authors: Pagani
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: malves@ioc.fiocruz.br.
authors: Marcelo Alves
authors: MA
authors: Ferreira
authors: Laboratório de Biotecnologia e Fisiologia de Infecções Virais, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: ajsg@ioc.fiocruz.br.
authors: Antonio S
authors: AS
authors: Gonçalves
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: pacorrea@ioc.fiocruz.br.
authors: Paloma Rezende
authors: PR
authors: Correa
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: wdegrave@fiocruz.br.
authors: Wim Maurits
authors: WM
authors: Degrave
authors: Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil. Electronic address: lmlima@ioc.fiocruz.br.
authors: Leila
authors: L
authors: Mendonça-Lima
mesh_terms: D001500:BCG Vaccine / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000187:drug effects* / Q000235:genetics; D015153:Blotting, Western; D004269:DNA, Bacterial / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000276:immunology; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009154:Mutation / Q000187:drug effects; D009163:Mycobacterium bovis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion; D014158:Transcription, Genetic / Q000187:drug effects*; D014376:Tuberculosis / Q000188:drug therapy
chemical_list: D001500:BCG Vaccine; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins
keywords: Gain of function mutation; Mycobacterium bovis BCG; TetR transcriptional regulator; Tuberculosis vaccine; rv3405c