At Last: Mesothele Silk

September 28, 2012

Tags: Mesothelae, Liphistius, Mygalomorphae, Hypochilus, Latrodectus, silk, proteins, evolution, Starrett, Garb, Kuelbs, Azubuike, Hayashi

I’ve been waiting for years to learn something about mesothele silk genes and silk proteins. The 90 or so species of mesothele make up the oldest surviving spider lineage. They now live only in Southeast Asia. As we explain in Spider Silk, mesotheles are sometimes considered living fossils (a term biologists don’t like nowadays; it gives the false impression that the organism hasn’t evolved at all since some distant point in the past) because they display characteristics so similar to the characteristics found in a 290-million-year-old fossil mesothele and so dissimilar from more “modern” spiders. For one thing, unlike other spiders, their abdomens are segmented, an outward display of spiders’ descent from segmented ancestors. If we want a full picture of spider silk evolution, mesothele silk genes and proteins are an obvious place to focus.

James Starrett, Jessica Garb, Amanda Kuelbs, Ugochi Azubuike, and Cheryl Hayashi have, for the first time, looked at the sequence of mesothele silk proteins by synthesizing cDNAs from the silk glands of Liphistius malayanus, a mesothele. They also looked at the sequences of silk proteins of six species of mygalomorph spider—these are the spiders in the second major spider lineage, which includes the tarantulas.

Neither mesotheles nor mygalomorphs spin the kinds of webs most non-arachnologists think of when we think “web.” They are incapable, for example, of spinning the kind of silk araneomorph spiders (the third major lineage) use as dragline or rapelling rope. (This largely accounts for silk researchers’ relative neglect of mesotheles and mygalomorphs: there’s no obvious industrial or other money-making application of this research). Instead, these spiders use silk to create linings for their burrows, or above-ground tubes such as pursewebs, or sheets that spread away from the spider’s hiding place to create a prey detector.

The research team also looked at the sequence of silk proteins produced by Hypochilus thorelli, a lampshade spider, one of the earliest-evolved still-surviving araneomorph spiders. Unlike the mesotheles and mygalomorphs, Hypochilus produces different kinds of silk strands specialized for different tasks. The researchers state that the motive for their study is the belief that “characterizing silk transcripts in mesotheles, mygalomorphs, and a basal araneomorph lineage allows for a better understanding of the evolutionary transition from substrate-borne, general-use silk fibers to aerial webs with task specific fibers spun by orb-weavers.” In other words, ancient spiders may have produced multiple silk proteins, but they seem to have used them interchangeably. Later in spider evolution, spiders use specific silks for specific jobs. Can silk protein sequences tell us how spiders got from there to here?

Previous to this research, two different classes of silk protein had been identified: spidroins, or spider fibroins, which make up the bulk of spider silk fibers, and egg case proteins (ECPs). ECPs were identified only in 2005 (whereas spidroins have been studied for decades) and only in Latrodectus hesperus, the Western black widow. (Given how few spider families have been sampled, though, it’s quite possible other spiders also produce them.)

Back in the early 1990s, Joachim Haupt and Jacqueline Kovoor conducted histology and histochemical studies on mesothele silk glands that showed that these spiders produced more than one kind of fibrous protein. What makes Starrett’s team’s findings so interesting is not just that they confirm Haupt and Kovoor’s study, but also that they found such a large variety of recognizable silk proteins in the mesothele: a spidroin plus six ECP-like proteins.

This appears to confirm the hypothesis that spidroins evolved very early in spider evolutionary history. Arachnologists have long assumed that the “original” spider produced just one spidroin, and that the genes scripting later-evolved spidroins probably evolved from the gene scripting that original spidroin. This study found just one spidroin in a mesothele, which makes that assumption seem more likely.

At the same time, the presence of the ECP-like proteins in addition to the spidroin shows that silk proteins started to diversify quite early in spider evolutionary history. These mesothele ECP-like proteins, like the mesothele spidroin, contain little repetition compared to the araneomorph black widow ECPs. But the selection pressures on mesothele silks are quite different from the selection pressures on black widow silks. Mesotheles live underground in burrows, and their eggs are laid and covered in silk underground. Mesothele silk proteins may be selected to cope with dampness or dangers such as mold or interfacing with soil or underground predators and parasites, whereas black widow silks may be selected to cope with sunlight, aridity, attack by above-ground predators and parasites, and wind forces, among other challenges.

The big question is: Why did these silk proteins start to diversify in the first place? It’s possible that mesotheles are not quite so indiscriminate in their use of different silk proteins as we think. In black widows, the ECPs knit with a specialized spidroin called tubuliform silk to form egg casing. Neither mesotheles nor mygalomorphs produce tubuliform silk. The research team believes that, because both ECPs and the ECP-like proteins are rich in the amino acid cysteine, and because cysteine plays a role in the knitting together of black widow ECPs and tubiliform spidroin, the mesothele ECP-like proteins may knit with that spider’s spidroin. Does the mesothele tailor its protein mix to lay down, for example, an egg blanket versus a trap door?

That isn’t an easy question to answer. Araneomorphs’ specialized silks are produced in specialized glands, and they can be observed as they emerge from distinct spinnerets. Mesothele glands don’t seem much different from each other. And closely observing a mesothele laying down silk requires unusual dedication: they undertake housekeeping chores in the dark, stop when exposed to light, and also, unlike mygalomorphs and araneomorphs, their spinnerets are inconveniently (for observers, not for the spiders) placed under their abdomens instead of exposed out at the end. So whether he or she chooses dissection and RNA extraction or focused observation or both, some researcher is going to have to contract an obsession with this question before we get an answer.

It’s also possible that the early diversification of these proteins is related to the different amounts of energy required for their synthesis. Spiders not entirely dependent on any one protein in their heavy dependence on silk may have been able to weather fluctuations in prey capture more easily.

What is clear is that this early diversification of silk proteins shows once again that silk genes are evolvable; that is, they have a propensity for evolution—they can duplicate and change slightly but still be useful. Eventually, under the right circumstances, accumulated changes may result in proteins suited to specific tasks.

The research team’s analysis of the mesothele spidroin, the newly found mygalomorph spidroins, the newly found Hypochilus spidroin, and previously known spidroins supports this scenario. They believe that their analysis shows that the common ancestor of the opisthotheles (the combined mygalomorphs and araneomorphs) had a minimum of five related spidroin genes, meaning that this gene duplication occurred before the mygalomorphs and araneomorphs went their separate ways. Mygalomorphs, like mesotheles, have pretty uniform silk glands. So this would mean that spidroins diversified before glands evolved to be specialized.

ECPs and ECP-like silk proteins are probably also much more diverse—it strains belief that they are produced only by mesotheles and black widows, which are pretty distant relatives. It’s much more likely that, due to the way the cDNA synthesis process works, it’s possible the team sampled their mygalomorph and Hypochilus specimens at a time when the genes scripting ECP-likes or ECPs weren’t being expressed. We’ll only know for sure when genome sequences—rather than just cDNA sequences—are available for various spiders. Then we’ll also be able to see what, if any, evolutionary connections there are between ECPs and spidroins.

Besides these basic findings, this study found a number of other interesting details that should be of interest to silk specialists. And like all good research, it points the way to a number of further research questions, which the authors point out: We know that the number of repeats has an influence on silk fiber formation and mechanical properties, but what influence does the length of repeats have? Do the silks of different mygalomorphs have different mechanical properties, and how are those properties related to the molecular architecture of their silk proteins? And finally, once the mechanical properties and functional properties of mesothele and different mygalomorph silks are understood, how have those properties been influenced by the natural selection pressures exerted by life underground?

Once you get tangled up in spider silk research, there’s really no end to its fascination.

Feet and Glue

October 7, 2011

Tags: silk, evolution, Theraphosidae, Rind, Gorb, Aphonopelma, Perez-Miles, Grammostola, Brachypelma, Peattie, Sahni, Latrodectus

[This post contains later amendments that may be instructive to other science writers. See the later posts on purported tarantula foot silk to see why.]

Three papers published this summer might at first seem unrelated. But read together, they pull the entire arc of spider silk evolution into sharper focus. Two papers indirectly address the evolutionary origins of spider silk production. The other demonstrates that the evolution of silk proteins has been central to spider evolution even after the extraordinary proliferation of silks that made the vertical orb web possible.

First, F. Claire Rind and colleagues reported that at least some tarantulas (which belong to the family Theraphosidae) do indeed secrete silk from their feet. This report appears to settle a controversy that first broke out in 2006, when a team of researchers led by Stanislav Gorb announced that they had persuaded a Costa Rica zebra tarantula, Aphonopelma seemanni, to walk on a nearly vertical surface covered with glass microscope slides. The researchers claimed that as the tarantula started to slip, it left behind “footprints” made up of miniscule silk fibers. If this observation held true, it could have important implications concerning the origin of spider silk production. Like all spiders, tarantulas secrete silk through abdominal spinnerets, small appendages ending in multiple spigots that are the outlets for the abdominal silk glands. Genetic studies have shown that spider spinnerets are the evolutionary descendants of the gill branches of ancient arthropod limbs. If spiders secreted silk from their limbs as well as through their spinnerets, this fact might not only cement the limb-spinneret connection but also suggest new hypotheses for the earliest origins and survival value of spider silk.

The Gorb report left some questions open, however. (more…)

Tough Genetic Stuff

October 18, 2010

Tags: genetics, protein structure, silk, Latrodectus, Zhao, Ayoub, Hayashi

Genetics papers can be pretty impenetrable to us non-geneticists. But geneticists don't write the way they do just so they'll be perceived as eggheads.

It took me, an English major with not much science background beyond high school courses, many months to learn how to decipher the genetics papers that inform Spider Silk. Of course, I was lucky enough to have Cay Craig guide me through these papers and steer me back on track when I veered astray. During this process, I came to realize that genes and genetic research is even more complicated than most of us non-biologists realize. For many of us, it's a mystery why news reports about exciting discoveries in genetics don't lead rapidly to successful medical or other practical applications.

I now get that it's no mystery, or conspiracy. This longer-than-usual post is an attempt to walk through an intriguing paper by a genetics team that writes unusually clearly. Even so, the paper is shot through with terms such as "paralog," "diploid," "retroposition," and "fluorescence in situ hybridization." These terms immediately convey images and lines of logic to other geneticists but gaping black holes to the rest of us. I'm going to avoid such terms as much as possible as I walk through the paper. But I think you'll still see how many interlocking and complex concepts and techniques evolutionary geneticists have to wrestle with, and why even dazzling genetics papers usually lead to more papers rather than to immediate, dramatic applications. (more…)




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