Endoplasmic Reticulum (ER) Membrane Proteins (LUNAPARKs) are Required for Proper Configuration of the Cortical ER Network in Plant Cells.

The endoplasmic reticulum (ER) is a large network made of membranous cisternae and tubules, which accounts for a large proportion of the total lipid bilayer endomembrane of the cell. In mammals and yeast, LUNAPARK proteins are preferentially localized at the three-way junctions of the ER network, stabilizing the junctions and establishing the ER architecture. We identified two Arabidopsis homologs and designated them LNPA and LNPB. Subcellular localization analysis with a non-dimerizable type of green fluorescent protein (GFP) revealed that both LNPA and LNPB are predominantly distributed throughout the ER, but not preferentially localized at the three-way junctions. Quantitative analysis of the network in the double mutant lnpa lnpb revealed that deficiency of LNPA and LNPB caused the cortical ER to develop poor ER cisternae and a less dense tubular network. These phenotypes are opposite to those of LNP-deficient mutants of yeast and mammals. Despite the importance of cysteine residues in the zinc finger motif of the yeast LNP homolog (Lnp1p), the corresponding cysteine residues of LNPA were not necessary for the stabilization of ER morphology because replacing the four cysteine residues in the zinc finger motif of the LNPA protein with alanine residues did not affect its function. A significant phenotype of lnpa lnpb is generation of large spherical structures from the ER. Formation of the structures might reduce the amounts of the ER membrane to be used for generating the network, resulting in poor development of the ER network. Taken together, our results suggest that plant LNPs function differently from those in yeast and mammals: they function to distribute ER membranes appropriately throughout the cells.