To bud where no bud has gone before

Acrobatic flexibility of the translocon T he machinery that inserts membrane proteins into the ER is unexpectedly fl exible, say Kida et al. Even with several segments of a multispanning membrane protein already looped around inside the pore, more can be inserted. Proteins are inserted into the ER in eukaryotes and the plasma membrane in prokaryotes by the Sec61-based translocon. Data from several studies, including crystal structures, indicate that the translo-con's pore consists of a single Sec61α subunit, thus creating a narrow channel. But the new studies suggest the pore is much larger than expected. The authors trapped intermediates in the translocation process by adding a streptavidin-binding peptide tag to the NH 2-terminal end of the inserted protein. When streptavidin was added, translocation stalled, resulting in intermediates. The addition of biotin removed the streptavidin and revived translocation. The intermediates revealed that stalled translocating peptides within the pore do not jam the translocon. Two distant hydrophilic segments of the same protein fi t within the pore at the same time. As each of these lipid-averse segments needs Sec61α to protect it from the membranous environment, the structure suggests that the pore is not as narrow as previously thought. Another intermediate revealed that a stalled hydrophilic domain did not prevent the subsequent membrane insertion of as many as six successive hydrophobic segments. To explain the fl exibility, the authors suggest that perhaps two Sec61 complexes—one for each hydrophilic segment—combine for a translocation event. This model is supported by previous EM images of Sec61 oligomers. It is also possible that a single Sec61 complex undergoes drastic, unpredicted conformational changes or is somehow made larger by accessory proteins. To bud where no bud has gone before L ike lightning, yeast bud sites never strike twice in the same place. Now, Tong et al. reveal that a zone of GTPase inhibition prevents a new bud site from overlapping with the previous site. The process of budding leaves behind a scar in the cell wall. Each cell cycle creates a new yeast scar, as new division sites never fall on top of previous ones. Scientists wondered whether the scars might make the cell wall too rigid for a new bud site to form there. But the new fi ndings show that the physical properties of the wall are not to blame. New bud sites repeatedly formed at the same site—on top of a scar—when a GTPase-activating …