Regulatory RNAs identification in response to iron availability and their mechanisms of action in Caulobacter crescentus

• Grant number: 15/07386-7
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): July 01, 2015
• Effective date (End): June 30, 2018
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Marilis Do Valle Marques
• Grantee: Laura Leaden
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 14/04046-8 - Regulatory networks of bacterial stress response, AP.TEM

Abstract

Small non-coding regulatory RNAs (sRNAs) may act on the translation of a target mRNA, favouring or preventing the formation of secondary structures, so that ribosomes may or not have free access to the RBS. Finally, sRNAs may change the mRNA stability, making it more stable or unstable. A global transcriptomic analysis performed in our laboratory identified C. crescentus genes whose expression was Fur-dependent in response to iron, but also a group of genes differentially expressed in response to iron independently of Fur. These results suggest the existence of a second regulator, probably a regulatory RNA as described for other organisms, and so far no sRNA involved in regulating iron homeostasis has been identified in this bacterium. This project proposes to identify small regulatory RNAs in iron starvation, through global RNA sequencing analysis (RNAseq). This approach allows the identification of differentially expressed genes with more sensitivity than DNA microarrays. Once identified, new RNAseq experiments will be carried out with a null mutant strain for that sRNA, to identifiy its post-transcriptional targets. Translational targets will be identified by mass spectrometry of protein extracts. Finally, the regulatory mechanisms will be determined using mutant strains for the ribonucleases RNase E, RNase R, RNase III, PNPase and Hfq.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LEADEN, LAURA; SILVA, LARISSA G.; RIBEIRO, RODOLFO A.; DOS SANTOS, NAARA M.; LORENZETTI, ALAN P. R.; ALEGRIA, THIAGO G. P.; SCHULZ, MARIANE L.; MEDEIROS, MARISA H. G.; KOIDE, TIE; MARQUES, V, MARILIS. Iron Deficiency Generates Oxidative Stress and Activation of the SOS Response in Caulobacter crescentus. FRONTIERS IN MICROBIOLOGY, v. 9, AUG 28 2018. (14/04046-8, 16/06378-3, 15/07386-7, 17/02127-9, 15/21038-1, 17/03052-2)

SILVA, LARISSA G.; LORENZETTI, ALAN P. R.; RIBEIRO, RODOLFO A.; ALVES, INGRID R.; LEADEN, LAURA; GALHARDO, RODRIGO S.; KOIDE, TIE; MARQUES, MARILIS V.. OxyR and the hydrogen peroxide stress response in Caulobacter crescentus. Gene, v. 700, p. 70-84, JUN 5 2019. (15/07386-7, 16/06378-3, 15/21038-1, 14/04046-8, 17/03052-2)

BALHESTEROS, HELOISE; SHIPELSKIY, YAN; LONG, NOAH J.; MAJUMDAR, ARITRI; KATZ, BENJAMIN B.; SANTOS, NAARA M.; LEADEN, LAURA; NEWTON, SALETE M.; MARQUES, MARILIS V.; KLEBBA, PHILLIP E.. TonB-Dependent Heme/Hemoglobin Utilization by Caulobacter crescentus HutA. Journal of Bacteriology, v. 199, n. 6, MAR 2017. (14/04046-8, 15/07386-7, 13/06873-6)