December 28 – Achieve. What’s the thing you most want to achieve next year? How do you imagine you’ll feel when you get it? Free? Happy? Complete? Blissful? Write that feeling down. Then, brainstorm 10 things you can do, or 10 new thoughts you can think, in order to experience that feeling today.
What do I want most? Get published three times in scholarly journals! Namely, in order that I envision the papers coming along, Molecular Microbiology, Journal of Biochemistry, and then finally Proceedings of the National Academy of Sciences.
How will I feel? Seriously? Is that a legitimate question here? I am going to feel fucking awesome!!!
Ten things I can do to get there? Oy vey…here comes the trouble.
- Figure out what alternative pathway hda seems to be mucking around with, either genetically or biochemically.
- Purify a bunch (more) Hda protein and demonstrate that it definitely impairs translesion synthesis DNA polymerases.
- Demonstrate that Hda is able to stabilize the replication fork and alter the ability for other accessory proteins to interact with the complex.
- Show biochemically that my mutant library is composed of both RIDA-competent and RIDA-incompetent mutants.
- Get a crystal structure of Hda, or a co-complex of Hda with the ß-subunit of DNA polymerase III.
- Selectively break interactions of Hda with other accessory proteins, and restore those interactions by compensatory mutations on said accessory proteins.
- Genetically demonstrate the ability to block and/or promote these interactions with mutants.
- Demonstrate the blatant redundancy of the initiation regulation pathways in cellular viability.
- Demonstrate that accessory functions of the initiation regulators were the evolutionary pressure for adoption as opposed to control of initiation.
- Make the grand corollary of what all this means in the greater scheme of breaking the microbial replication cycle and propose a new, unique pathway of antimicrobial targets.
Nobody said I couldn’t be dreaming when I thought all of this up. But it’s an entirely possible order if the experiments go in my favor…