The YopD translocator of Yersinia pseudotuberculosis is a multifunctional protein comprised of discrete domains.
To establish an infection, Yersinia pseudotuberculosis utilizes a plasmid-encoded type III translocon to microinject several anti-host Yop effectors into the cytosol of target eukaryotic cells. YopD has been implicated in several key steps during Yop effector translocation, including maintenance of yop regulatory control and pore formation in the target cell membrane through which effectors traverse. These functions are mediated, in part, by an interaction with the cognate chaperone, LcrH. To gain insight into the complex molecular mechanisms of YopD function, we performed a systematic mutagenesis study to search for discrete functional domains. We highlighted amino acids beyond the first three N-terminal residues that are dispensable for YopD secretion and confirmed that an interaction between YopD and LcrH is essential for maintenance of yop regulatory control. In addition, discrete domains within YopD that are essential for both pore formation and translocation of Yop effectors were identified. Significantly, other domains were found to be important for effector microinjection but not for pore formation. Therefore, YopD is clearly essential for several discrete steps during efficient Yop effector translocation. Recognition of this modular YopD domain structure provides important insights into the function of YopD.
Olsson
Jan
J
Department of Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden.
Edqvist
Petra J
PJ
Bröms
Jeanette E
JE
Forsberg
Ake
A
Wolf-Watz
Hans
H
Francis
Matthew S
MS
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D001426:Bacterial Proteins / Q000502:physiology; D001483:Base Sequence; D006367:HeLa Cells; D006461:Hemolysis; D006801:Humans; D018832:Molecular Chaperones / Q000502:physiology; D008969:Molecular Sequence Data; D021381:Protein Transport; D015011:Yersinia pseudotuberculosis / Q000502:physiology*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D018832:Molecular Chaperones; C118339:SycD protein, bacteria; C078910:YopD protein, Yersinia