Group (ethan itovitch)
For the htgaa final project groups are tasked with finding solutions for making the MS2 bacteriophage more effective against its target: E. Coli. This is of interest because over the years we have over used antibiotics and in the process have created superbugs: Bacteria that are resistant to antibiotics. It is estimated that superbugs will be responsible for more deaths per year than cancer by 2050.
Using bacteriophages to target bacteria is of interest because they are highly specific to their target and offer an alternative to using antibiotics which kills everything (including good bacteria)
That being said, bacteria have defences against their phages: Evolution. Bacteria are able to mutate in a constant arms race against their phages and avoid being exterminated.
Our goal in this project is to find ways to modify the MS2 DNA so that it can have a head start against E Coli.
Ms2 is a tiny bacteriophage with a single-stranded RNA genome. It enters it’s RNA inside E. Coli. through it’s F-pilin. once inside, it translates the RNA to create new copies of itself. Finally a lysis protein is expressed and lyses the bacteria allowing the MS2 phage to be released and infect more targets.
The Lysis protein is crucial to the MS2 phages reproduction cycle. However, it is heavily dependant on E. Coli’s DnaJ chaperone protein and one of the defences that E. Coli. can employ to resist against the MS2 phage is to mutate this protein. It is commonly understood that the DnaJ’s role is to help the Lysis protein with proper protein folding since it is a chaperone protein after all. However, I think it might serve a different purpose.
In the following paper: “MS2 Lysis of Escherichia coli Depends on Host Chaperone DnaJ”: https://pmc.ncbi.nlm.nih.gov/articles/PMC5446614/ it is stated that the lysis protein has 2 components, the N terminus (the squiggly line in the image) which is what binds to the DnaJ & the C terminus which is what enters the cell membrane and actually does the damage lysing the membrane. Additionally it is stated that when the N terminus section was removed the problem wasn’t that the protein didn’t fold correctly, it was that lysing occurred too quickly.
If this is the case, then maybe the interaction with DnaJ isn’t for protein folding but for delaying the lysis protein from making it to the membrane giving the phage more time to prepare for the lysis. This would explain how in the absence of the N terminus, lysing occurs too quick.
sequence:
METRFPQQSQQTPASTNRRRPFKHEDYPCRRQQRSSTLYVLIFLAIFLSKFTNQLLLSLLEAVIRTVTTLQQLLT