New & Noteworthy

The Baby Bear of Proteins

July 31, 2013

In the story of Goldilocks and the Three Bears, Goldilocks always likes Baby Bear’s stuff best. Baby Bear has the most comfortable bed, the best porridge, and so on.

The Three Bears

When it comes to protein levels, yeast cells can be just as picky as Goldilocks.

The reason Goldilocks likes Baby Bear’s things are that they are just right. They are neither too hard nor too soft, too hot nor too cold, too big nor too little.

Turns out that when it comes to certain proteins, the yeast S. cerevisiae is sort of like Goldilocks…it likes to have them at just the right levels. Too much or too little protein can throw things out of whack.

This idea is supported in a new study in GENETICS, where Sasanuma and coworkers find that a key helicase in yeast, Srs2p, needs to be present in just the right amounts for meiotic recombination to go off without a hitch. In particular, they show that this protein affects meiotic recombination by interfering with the assembly of filaments containing another protein, Rad51p.

Meiotic recombination starts off with Spo11p making a double stranded (DS) break in the DNA. This DS DNA is then trimmed back so that there is a 3′ overhang of single stranded DNA which is then coated with replication protein A (RPA), Rad51p, and Dmc1p. The coated single stranded DNA then invades a stretch of homologous DNA and recombination can begin. 

One of the first things Sasanuma and coworkers did was to show that toying with Srs2p levels has a negative effect on meiosis in general. Too little Srs2p brings spore viability down to 36.8% of wild type, and overexpressing it brings spore viability down to 22.4%.  Clearly Srs2p is a bit like Goldilocks…the amount has to be just right.

The authors next set out to determine how overexpressing Srs2p affects meiosis so profoundly. They showed that too much Srs2p delays the start of meiosis, causes chromosomes to end up in the wrong places, and stunts the repair of DS DNA breaks. Basically, extra Srs2p inhibits meiotic recombination.

They next looked at areas on the DNA where both Rad51p AND Dcm1p were bound, and found that too much Srs2p keeps Rad51p but not Dcm1p off the DNA. When either of these proteins binds to DNA, it forms foci that are visible as dots when the proteins are detected with fluorescent antibodies. While a wild type strain had roughly equal numbers of Dcm1p and Rad51p foci, there were four fold fewer Rad51p foci when Srs2p was overexpressed. Clearly Srs2p was keeping Rad51p-DNA complexes from forming. 

Srs2p can act as a translocase, and it can also bind Rad51p. Sasanuma and coworkers asked which of these functions is essential to its ability to disassemble Rad51p filaments on DNA. Using srs2 mutants that were blocked in just one of these functions, they showed that the translocase mutant was completely unable to remove Rad51p from DNA during meiosis. The Rad51p-binding mutant could still cause Rad51p to dissociate from chromosomes, although at a reduced rate compared to wild type. So the translocase activity is essential, while Rad51p binding is not.

Although it was known that in vitro Srs2p can cause Rad51p-DNA filaments to disassemble, this study is the first to establish that it actually happens in vivo during meiosis. The requirement for the translocase activity suggests that Srs2p may actually move along the filaments as it disassembles them. And this work also shows that just like Goldilocks with her bowl of porridge, the cell needs an amount of Srs2p that is not too big, not too little, but just right.