I think your results clearly show that transduction isn't a preferred mechanism of transfer for staphylococcal plasmids! :p

Microbial Culture Club, Schott Duran Duran, Fleetwood MacConkey

Microbial Culture Club, Schott Duran Duran, Fleetwood MacConkey

We certainly haven’t finished with RdfS yet. Our molecular genetics data clearly suggest RdfS has a more direct role in the activation of ICE conjugation genes, we just don’t know exactly how yet. Hopefully it won't take another 20 years to find out!

We propose that RdfS oligomers recognise larger DNA sites, perhaps as preformed oligomers and through an indirect readout mechanism that facilitates binding specificity over large regions with low sequence conservation.

Callum identified several large 30-40bp DNA regions specifically bound by RdfS but with very little sequence conservation. He also showed the unique N-terminal helix was critical for ICE excision and conjugation.

wHTH domains are common and the RdfS structure could been boring. Thankfully it wasn't. RdfS in the absence of DNA forms infinite helical polymers joined together by a unique N-terminal helix that absent in other wHTH excisionases.

Much molecular genetics and 20 years later, Callum and Charlie Bond use some very clever solutions to solve the RdfS structure from an unusual crystal doi.org/10.1107/S205... demonstrating that like other excisionases, RdfS is a unique winged-helix-turn-helix protein.

Inspired by undergrad bioinformatics lectures (P. Stockwell & C. Brown), I used PSI-BLASTP to iteratively search for excisionase sequences in rhizobia, and identified rdfS as the first gene in an operon containing transfer genes and a conjugative relaxase RlxS. doi.org/10.1111/j.13...

I was fascinated by phage integrases and was sure the tyrosine recombinase IntS, of the symbiosis ICE, must have an excisionase to stimulate excision. But unlike most integrases, intS didn’t have an excisionase gene nearby.

This is a special paper for me, as rdfS was the first gene I ever made a functional prediction for 20 years ago as a 1st-year PhD student. The symbiosis island had recently been discovered (Ronson laboratory) and the 500kb sequence had just been completed (Sanger sequencing!).

Thanks!

Is it possible to share a copy of this? I don't have access but would love to read!

I personally don't agree. I think more thorough review is more important, even when it doesn't go your way. Also, the pressure to publish so frequently somewhat diminishes the value of individual publications and supports the publish or perish situation. I prefer to spend more time on publications.

Could you add me please and @mgeaus.bsky.social ?