In situ structures of secretins from bacterial type II secretion system reveal their membrane interactions and...

The GspD secretin is the outer membrane channel of the bacterial type II secretion system (T2SS) which secrets diverse effector proteins or toxins that cause severe diseases such as diarrhea and cholera. GspD needs to translocate from the inner to the outer membrane to exert its function, and this process is an essential step for T2SS to assemble. Here, we investigate two types of secretins discovered so far in Escherichia coli , GspD _α and GspD _β , respectively. By electron cryotomography subtomogram averaging, we determine in situ structures of all the key intermediate states of GspD _α and GspD _β in the translocation process, with resolution ranging from 9 Å to 19 Å. In our results, GspD _α and GspD _β present entirely different membrane interaction patterns and ways of going across the peptidoglycan layer. We propose two distinct models for the membrane translocation of GspD _α and GspD _β , providing a comprehensive perspective on the inner to outer membrane biogenesis of T2SS secretins.