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Signaling in a Crowd

February 18, 2014

Like a lonely “secrete-and-sense” cell, this skier can only encourage himself.

There are two very different kinds of sports in the Winter Olympics (and in all sporting competitions really).  In one set, it is the athletes alone out on the ice or sliding down the slope, trying to get the best time they can.  They can only use themselves as the motivator.

In another set of sports, like speed skating, athletes compete directly with one another.  Here they can use each other to push themselves to go faster, farther, etc.

The key to each is obviously the proximity of other athletes.  If there are a bunch of athletes around you, you will all do better by feeding off each other’s signals.  If you are by yourself, then only you can produce the signals to motivate yourself to go faster.

Youk and Lim show in a new study that the same sort of thing happens in cells that can both secrete and sense the same signal.  If there aren’t a lot of cells around they tend to signal themselves, but in a crowded place, they are all signaling each other. 

This may seem a bit esoteric but it really isn’t.  These sorts of “secrete-and-sense” systems are common in biology.  Cell types from bacteria to our own T cells have them, and they allow for a surprisingly wide range of responses.  Understanding how these systems work will explain a lot of biology and, perhaps, help scientists create new sensing systems for bioengineered beasts.

Youk and Lim used our favorite organism Saccharomyces cerevisiae to study this widespread signaling system.  They created a bevy of strains that can either secrete and sense alpha factor or that can only sense the pheromone.  They grew varieties of these two strains together under various conditions to determine when the “secrete-and-sense” strains could also signal to the “sense only” strains.  Like our athletes, the cell concentration was important.  But so too were the levels of alpha factor and receptor.

The authors first created a strain that senses the presence of alpha factor with the Ste2p receptor and in response turns on GFP through the FUS1 promoter.  (The strain is deleted for FAR1 to prevent cell cycle arrest.) As expected, increasing amounts of alpha factor resulted in increased levels of GFP.

It is from this strain they created their “secrete-and-sense” and “sense only” strains.  The “secrete-and-sense” strain included a doxycycline inducible promoter driving the alpha factor gene.  The more doxycycline, the more alpha factor it makes, resulting in more GFP.  To tell the two strains apart in experiments, they added a second reporter, mCherry, under a constitutive promoter to the “sense only” strain.  Now in their experiments they can distinguish between the strains that glow only green and those that glow red and, sometimes, green.

The first experiment was simply to see what effect differing cell and alpha factor concentrations had on the two strains’ ability to glow green.  At low cell and doxycycline concentrations, only the “secrete-and-sense” strain glowed green.  This makes sense, as too little alpha factor was made to get to the relatively distant neighbors.  At high cell and doxycycline concentrations, both glowed green almost indistinguishably.  Here the system was flooded with enough alpha factor for everyone to respond.

The results were less binary at either low cell and high doxycycline concentrations or high cell and low doxycycline concentrations.  Under either of these conditions, the “sense only” strain did glow green although at a much slower rate.

Youk and Kim didn’t stop there.  They also tested whether the amount of receptor affected these results.  When the two strains expressed high levels of receptor, the amount of alpha factor didn’t matter at low cell concentrations—only the “secrete-and-sense” strain glowed green.  This makes sense as the strain can quickly suck up any amount of alpha factor it makes.  Again at high cell concentrations the differences disappear.

In a final set of experiments the authors created positive feedback loops and signal degradation systems, which are both very common in nature.  The positive feedback loop was created by putting the doxycycline activator, rtTA, under the control of doxycycline, and a signal degradation system was engineered using Bar1p, a protease that degrades alpha factor.  Using these systems they were able to show that at low cell concentration, low Bar1p expression, and strong positive feedback, individual cells were either on or off.  This sort of activity may be important in nature, where under certain conditions a response may be beneficial and in others a response may not.  This bet hedging means that the population can survive under both sets of conditions.

It is amazing that such a simple set of conditions can lead to so many different responses, almost as varied as the performances of Olympic athletes.  These findings not only help to explain how these deceptively simple systems work and why they are so common in nature, but might also be incredibly useful in setting up synthetic secrete-and-sense circuits for biotechnology applications.  

by D. Barry Starr, Ph.D., Director of Outreach Activities, Stanford Genetics