SKP1

A team of researchers reconstructed long-extinct proteins of a UV protection system of cyanobacteria. The surprising result: the proteins were already compatible with each other when they first met. This discovery expands the knowledge horizon on the rules of evolution. Proteins are the key players for virtually all molecular processes within the cell. To fulfil their diverse functions, they have to interact with other proteins. Such protein-protein interactions are mediated by highly complementary surfaces, which typically involve many amino acids that are positioned precisely to produce a tight, specific fit between two proteins. However, comparatively little is known about how such interactions are created during evolution. Classical evolutionary theory suggests that any new biological feature involving many components (like the amino acids that enable an interaction between proteins) evolves in a stepwise manner. According to this concept, each tiny functional improvement is driven by the power of natural selection because there is some benefit associated with the feature. However, whether protein-protein interactions also always follow this trajectory was not entirely known. Using a highly interdisciplinary approach, an international team led by Max Planck researcher Georg Hochberg from the Terrestrial Microbiology in Marburg have now shed new light on this question. Their study provides definitive evidence that highly complementary and biologically relevant protein-protein interactions can evolve entirely by chance. Proteins cooperate in a photoprotection system The research team made their discovery in a biochemical system that microbes use to adapt to stressful light conditions. Cyanobacteria use sunlight to produce their own food through photosynthesis. Since much light damages the cell, cyanobacteria have evolved a mechanism known as photoprotection: if light intensities become dangerously high, a light intensity sensor named Orange Carotenoid Protein (OCP) changes its shape. In this activated form, OCP protects the cell by converting excess light energy into harmless heat. In order to return into its original state, some OCPs depend on a second protein: The Fluorescence Recovery Protein (FRP) binds to activated OCP1 and strongly accelerates its recovery. 'Our question was: Is it possible that the surfaces that allow these two proteins to form a complex evolved entirely by accident, rather than through direct natural selection?' says Georg Hochberg. 'The difficulty is that the end result of both processes looks the same, so we usually cannot tell why the amino acids required for some interaction evolved -- through natural selection for the interaction or by chance. To tell them apart, we would need a time machine to witness the exact moment in history these mutations occurred, 'Georg Hochberg explains. Luckily, recent breakthroughs in molecular and computational biology has equipped Georg Hochberg and his team with a laboratory kind of time machine: ancestral sequence reconstruction. In addition, the light protection system of cyanobacteria, which is under study in the group of Thomas Friedrich from Technische Universität Berlin since many years, is ideal for studying the evolutionary encounter of two protein components. Early cyanobacteria acquired the FRP proteins from a proteobacterium by horizontal gene transfer. The latter had no photosynthetic capacity itself and did not possess the OCP protein. To work out how the interaction between OCP1 and FRP evolved, graduate student Niklas Steube inferred the sequences of ancient OCPs and FRPs that existed billions of years ago in the past, and then resurrected these in the laboratory. After translation of the amino acid sequences into DNA he produced them using E. coli bacterial cells in order to be able to study their molecular properties. A fortunate coincidence The Berlin team then tested whether ancient molecules could form an interaction. This way the scientists could retrace how both protein partners got to know each other. 'Surprisingly, the FRP from the proteobacteria already matched the ancestral OCP of the cyanobacteria, before gene transfer had even taken place. The mutual compatibility of FRP and OCP has thus evolved completely independently of each other in different species, says Thomas Friedrich. This allowed the team to prove that their ability to interact must have been a happy accident: selection could not plausibly have shaped the two proteins' surfaces to enable an interaction if they had never met each other. This finally proved that such interactions can evolve entirely without direct selective pressure. 'This may seem like an extraordinary coincidence,' Niklas Steube says. 'Imagine an alien spaceship landed on earth and we found that it contained plug-shaped objects that perfectly fit into human-made sockets. But despite the perceived improbability, such coincidences could be relatively common. But in fact, proteins often encounter a large number of new potential interaction partners when localisation or expression patterns change within the cell, or when new proteins enter the cell through horizontal gene transfer.' Georg Hochberg adds, 'Even if only a small fraction of such encounters ends up being productive, fortuitous compatibility may be the basis of a significant fraction of all interactions we see inside cells today. Thus, as in human partnerships, a good evolutionary match could be the result of a chance meeting of two already compatible partners.'

Researchers have helped reveal the most detailed picture to date of important biological 'antennae.' Michigan State University researchers and colleagues at the University of California Berkeley, the University of South Bohemia and Lawrence Berkeley National Laboratory have helped reveal the most detailed picture to date of important biological "antennae." Nature has evolved these structures to harness the sun's energy through photosynthesis, but these sunlight receivers don't belong to plants. They're found in microbes known as cyanobacteria, the evolutionary descendants of the first organisms on Earth capable of taking sunlight, water and carbon dioxide and turning them into sugars and oxygen. Published Aug. 31 in the journal Nature, the findings immediately shed new light on microbial photosynthesis -- specifically, how light energy is captured and sent to where it's needed to power the conversion of carbon dioxide into sugars. Going forward, the insights could also help researchers remediate harmful bacteria in the environment, develop artificial photosynthetic systems for renewable energy and enlist microbes in sustainable manufacturing that starts with the raw materials of carbon dioxide and sunlight. "There's a lot of interest in using cyanobacteria as solar-powered factories that capture sunlight and convert it into a kind of energy that can be used to make important products," said Cheryl Kerfeld, Hannah Distinguished Professor of structural bioengineering in the College of Natural Science. "With a blueprint like the one we've provided in this study, you can start thinking about tuning and optimizing the light-harvesting component of photosynthesis." "Once you see how something works, you have a better idea of how you can modify it and manipulate it. That's a big advantage," said Markus Sutter, a senior research associate in the Kerfeld Lab, which operates at MSU and Berkeley Lab in California. The cyanobacterial antenna structures, which are called phycobilisomes, are complex collections of pigments and proteins, which assemble into relatively massive complexes. For decades, researchers have been working to visualize the different building blocks of phycobilisomes to try to understand how they're put together. Phycobilisomes are fragile, necessitating this piecemeal approach. Historically, researchers have been unable to get the high-resolution images of intact antennae needed to understand how they capture and conduct light energy. Now, thanks to an international team of experts and advances in a technique known as cryo-electron microscopy, the structure of a cyanobacterial light harvesting antenna is available with nearly atomic resolution. The team included researchers from MSU, Berkeley Lab, the University of California, Berkeley and the University of South Bohemia in the Czech Republic. "We were fortunate to be a team made up of people with complementary expertise, people who worked well together," said Kerfeld, who is also a member of the MSU-DOE Plant Research Laboratory, which is supported by the U.S. Department of Energy. "The group had the right chemistry." 'A long journey full of nice surprises' "This work is a breakthrough in the field of photosynthesis," said Paul Sauer, a postdoctoral researcher in Professor Eva Nogales' cryogenic electron microscopy lab at Berkeley Lab and UC Berkeley. "The complete light-harvesting structure of a cyanobacteria's antenna has been missing until now," Sauer said. "Our discovery helps us understand how evolution came up with ways to turn carbon dioxide and light into oxygen and sugar in bacteria, long before any plants existed on our planet." Along with Kerfeld, Sauer is a corresponding author of the new article. The team documented several notable results, including finding a new phycobilisome protein and observing two new ways that the phycobilisome orients its light-capturing rods that hadn't been resolved before. "It's 12 pages of discoveries," said María Agustina Domínguez-Martín of the Nature report. As a postdoctoral researcher in the Kerfeld Lab, Domínguez-Martín initiated the study at MSU and brought it to completion at the Berkeley Lab. She is currently at the University of Cordoba in Spain as part of the Marie Sk?owdoska-Curie Postdoctoral Fellowship. "It's been a long journey full of nice surprises." One surprise, for example, came in how a relatively small protein can act as a surge protector for the massive antenna. Before this work, researchers knew the phycobilisome could corral molecules called orange carotenoid proteins, or OCPs, when the phycobilisome had absorbed too much sunlight. The OCPs release the excess energy as heat, protecting a cyanobacterium's photosynthetic system from burning up. Until now, there's been debate about how many OCPs the phycobilisome could bind and where those binding sites were. The new research answers these fundamental questions and offers potentially practical insights. This kind of surge-protecting system -- which is called photoprotection and has analogs in the plant world -- naturally tends to be wasteful. Cyanobacteria are slow to turn their photoprotection off after it has done its job. Now, with the complete picture of how the surge protector works, researchers can design ways to engineer "smart," less wasteful photoprotection, Kerfeld said. And, despite helping make the planet habitable for humans and countless other organisms that need oxygen to survive, cyanobacteria have a dark side. Cyanobacteria blooms in lakes, ponds and reservoirs can produce toxins that are deadly to native ecosystems as well as humans and their pets. Having a blueprint of how the bacteria not only collect the sun's energy, but also protect themselves from too much of it could inspire new ideas to attack harmful blooms. Beyond the new answers and potential applications this work offers, the researchers are also excited about the new questions it raises and the research it could inspire. "If you think of this like Legos, you can keep building up, right? The proteins and pigments are like blocks making the phycobilisome, but then that's part of the photosystem, which is in the cell membrane, which is part of the entire cell," Sutter said. "We're climbing up the ladder of scale in a way. We've found something new on our rung, but we can't say we've got the system settled." "We've answered some questions, but we've opened the doors on others and, to me, that's what makes it a breakthrough," Domínguez-Martín said. "I'm excited to see how the field develops from here." This work was supported by the U.S. Department of Energy Office of Science, the National Institutes of Health, the Czech Science Foundation and the European Union's Horizon 2020 research and innovation program.