Abstract

Attachment of glycosylphosphatidylinositols (GPIs) to the C-termini of proteins is one of the most common posttranslational modifications in eukaryotic cells. GPI8/PIG-K is the catalytic subunit of the GPI transamidase complex catalyzing the transfer en bloc GPI to proteins. In this study, a T-DNA insertional mutant of rice with temperature-dependent drooping and fragile (df) shoots phenotype was isolated. The insertion site of the T-DNA fragment was 879 bp downstream of the stop codon of the OsGPI8 gene, which caused introns retention in the gene transcripts, especially at higher temperatures. A complementation test confirmed that this change in the OsGPI8 transcripts was responsible for the mutant phenotype. Compared to control plants, internodes of the df mutant showed a thinner shell with a reduced cell number in the transverse direction, and an inhomogeneous secondary wall layer in bundle sheath cells, while many sclerenchyma cells at the tops of the main veins of df leaves were shrunken and their walls were thinner. The df plants also displayed a major reduction in cellulose and lignin content in both culms and leaves. Our data indicate that GPI anchor proteins play important roles in biosynthesis and accumulation of cell wall material, cell shape, and cell division in rice.

Highlights

  • Attachment of glycosylphosphatidylinositol (GPI) moieties to protein C-termini is one of the most common posttranslational modifications in eukaryotic cells [1]

  • Intron retention and alternative splicing are associated with polyadenylation sites in plants [31]

  • Like Arabidopsis [12], rice has homologs of all five subunits, with all but one subunit encoded by only one gene: they are LOC_Os02g12740, which is a homolog of At1g08750 and Gpi8/PIG-K, LOC_Os01g48980 (At5g19130, Gaa1p/GAA1), LOC_Os01g67960 (At3g07180, Gpi17p/PIG-S), LOC_Os11g28980 (At3g07140, Gpi16p/PIG-T), LOC_Os02g46350, and LOC_Os01g64990 (At1g12730 and At3g27325, Gab1p/PIG-U)

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Summary

Introduction

Attachment of glycosylphosphatidylinositol (GPI) moieties to protein C-termini is one of the most common posttranslational modifications in eukaryotic cells [1]. Proteins modified by GPI attachment are called GPI anchor proteins (GPI-APs). The mature GPIs are transferred en bloc to proteins by GPI transamidase (GPI-T) complexes in the ER. GPI8 and GPAA1 are the catalytic subunits of GPI-T [3]. GPAA1 catalyzes the formation of a bond between the substrate protein’s omega-site and the GPI lipid anchor’s phosphoethanolamine [4]. GPI8 has a dual role: it carries out the proteolytic cleavage of the C-terminal signal sequence of the precursor protein, forms an amide bond between the protein and the ethanolamine phosphate of the GPI [5]

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