Adding a new member to the CadR regulatory network in Pseudomonas aeruginosa using REPSA and bioinformatics
Disciplines
Biochemistry | Molecular Biology
Abstract (300 words maximum)
Pseudomonas aeruginosa is a common bacterium that can infect human hosts. Currently, P. aeruginosa is classified as a "critical priority pathogen" by the World Health Organization due to its resistance to multiple antibiotics and being one of the leading causes of nosocomial infections - infections that come about during health care that were not present to begin with. One way researchers study the basic biology of pathogenic bacteria is by understanding how the bacteria control gene expression. Deciphering how genes are turned on or off in response to environmental stimuli can often lead to the development of improved treatment strategies. Transcription factors are proteins that bind to a specific DNA sequence to regulate gene expression. They recognize and react to factors in their environment to know when to and when not to bind DNA. The CadR – cadmium regulator – transcription factor is found in P. aeruginosa genome and activates the transcription of CadA, a cadmium exporter. CadR has been found to be responsive to both cadmium, which is not essential for cells, and zinc, which is essential for cells. Since zinc often plays a fundamental role in bacterial pathogenicity, we sought to understand the complete regulatory network for the CadR transcription factor. Using an iterative selection technique, Restriction Endonuclease Protection Selection and Amplification (REPSA), the DNA binding sequence for CadR was discovered. Mapping this sequence to the P. aeruginosa genome, we identified a genomic region that had not been cited as being regulated by CadR.Through bioinformatic approaches, we were able to determinethat this binding sequence was in the promoter region of apreviously uncharacterized, Zinc Ribbon Domain-containingprotein product. Through in vivo experimentation, we validatedthat this novel gene product is in the regulatory network ofCadR.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Michael Van Dyke, PhD
Additional Faculty
John Barrows, Biochemistry and Biology,
jbarro51@kennesaw.edu
Adding a new member to the CadR regulatory network in Pseudomonas aeruginosa using REPSA and bioinformatics
Pseudomonas aeruginosa is a common bacterium that can infect human hosts. Currently, P. aeruginosa is classified as a "critical priority pathogen" by the World Health Organization due to its resistance to multiple antibiotics and being one of the leading causes of nosocomial infections - infections that come about during health care that were not present to begin with. One way researchers study the basic biology of pathogenic bacteria is by understanding how the bacteria control gene expression. Deciphering how genes are turned on or off in response to environmental stimuli can often lead to the development of improved treatment strategies. Transcription factors are proteins that bind to a specific DNA sequence to regulate gene expression. They recognize and react to factors in their environment to know when to and when not to bind DNA. The CadR – cadmium regulator – transcription factor is found in P. aeruginosa genome and activates the transcription of CadA, a cadmium exporter. CadR has been found to be responsive to both cadmium, which is not essential for cells, and zinc, which is essential for cells. Since zinc often plays a fundamental role in bacterial pathogenicity, we sought to understand the complete regulatory network for the CadR transcription factor. Using an iterative selection technique, Restriction Endonuclease Protection Selection and Amplification (REPSA), the DNA binding sequence for CadR was discovered. Mapping this sequence to the P. aeruginosa genome, we identified a genomic region that had not been cited as being regulated by CadR.Through bioinformatic approaches, we were able to determinethat this binding sequence was in the promoter region of apreviously uncharacterized, Zinc Ribbon Domain-containingprotein product. Through in vivo experimentation, we validatedthat this novel gene product is in the regulatory network ofCadR.