Sunday, August 24, 2014

I don't even know what to think about this.

I've been recently thinking hard about how hfq influences competence and I'm stumped. Here are the facts:

Removing hfq in KW20
- Competence is lowered 10 fold when cells are in MIV
- Competence fails to increase as cells become dense (transformation frequency remains <5E-6)
- Adding cAMP temporarily increases competence back to expected wild type + cAMP levels but drops off again quickly

Removing hfq in sxy-1
- Competence in MIV is pretty much the same between the sxy-1 mutant and the double mutant
- Competence reaches MIV levels when growing in log phase (>10x higher than single mutant sxy-1)
- Double mutant in general is more competent than single mutant
- When adding cAMP cells appear to become maximally competent with about 1 in 100 cells transformed

Apparently adding a mutation that lowers competence in wild type to a hypercompetent mutant with a single point mutation increases competence even more so? I don't get it. As if it didn't get confusing enough....

Removing hfq in sxy-2
- Competence decreases about 5 fold for the double mutant in MIV
- Competence is a little lower for the double mutant in log phase (~1E-4) and fails to increase as the cells become more dense
- Not sure about the effect of cAMP yet.

The mutations in sxy-1 and sxy-2 are literally under five bases apart and when added to the wild type background, have a similar phenotype. Yet when we remove hfq from the sxy-1 mutant we see increased competence under almost every condition and when we remove hfq from the sxy-2 mutant, we see decreased competence. I think this is strong evidence that hfq may directly interact with the sxy mRNA in some capacity: the question is how?