Role of vesicle-inducing protein in plastids 1 in cpTat transport at the thylakoid.

VIPP1 has been shown to be required for the proper formation of thylakoid membranes. However, studies on VIPP1 itself, as well as on PspA, its bacterial homolog, suggests that this protein may be involved in a number of additional functions, including protein translocation. The role of VIPP1 in protein translocation in the chloroplast has not been investigated. To this end, we conducted in vitro thylakoid protein transport assays to look at the effect of VIPP1 on the cpTat pathway, which is one of three translocation pathways found in both the chloroplast and its bacterial progenitor. We found that VIPP1 does indeed enhance protein transport through the cpTat pathway by up to 100%. The VIPP1 effect on cpTat activity occurs without interacting with the substrates or components of the translocon, and does not alter the energy potentials driving this translocation pathway. Instead, VIPP1 greatly enhances the amount of substrate bound productively to the thylakoids. Moreover, the presence of increasing VIPP1 concentrations in the reactions resulted in greater interactions between thylakoid membranes. Taken together, these results demonstrate a stimulatory role for VIPP1 in cpTat transport by enhancement of substrate binding, probably to the membrane lipid regions of the thylakoid. We propose a model in which VIPP1 facilitates reorganization of the thylakoid structure to increase substrate access to productive binding regions of the membrane as an early step in the cpTat pathway.