title: Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis.
abstract: Delta-1-pyrroline-5-carboxylate synthetase enzymes, which catalyse the rate-limiting step of proline biosynthesis, are encoded by two closely related P5CS genes in Arabidopsis. Transcription of the P5CS genes is differentially regulated by drought, salinity and abscisic acid, suggesting that these genes play specific roles in the control of proline biosynthesis. Here we describe the genetic characterization of p5cs insertion mutants, which indicates that P5CS1 is required for proline accumulation under osmotic stress. Knockout mutations of P5CS1 result in the reduction of stress-induced proline synthesis, hypersensitivity to salt stress, and accumulation of reactive oxygen species. By contrast, p5cs2 mutations cause embryo abortion during late stages of seed development. The desiccation sensitivity of p5cs2 embryos does not reflect differential control of transcription, as both P5CS mRNAs are detectable throughout embryonic development. Cellular localization studies with P5CS-GFP gene fusions indicate that P5CS1 is sequestered into subcellular bodies in embryonic cells, where P5CS2 is dominantly cytoplasmic. Although proline feeding rescues the viability of mutant embryos, p5cs2 seedlings undergo aberrant development and fail to produce fertile plants even when grown on proline. In seedlings, specific expression of P5CS2-GFP is seen in leaf primordia where P5CS1-GFP levels are very low, and P5CS2-GFP also shows a distinct cell-type-specific and subcellular localization pattern compared to P5CS1-GFP in root tips, leaves and flower organs. These data demonstrate that the Arabidopsis P5CS enzymes perform non-redundant functions, and that P5CS1 is insufficient for compensation of developmental defects caused by inactivation of P5CS2.
authors.affiliation: Institute of Plant Biology, Biological Research Center, H-6726 Szeged, Hungary.
authors.forename: Gyöngyi
authors.initials: G
authors.lastname: Székely
authors.forename: Edit
authors.initials: E
authors.lastname: Abrahám
authors.forename: Agnes
authors.initials: A
authors.lastname: Cséplo
authors.forename: Gábor
authors.initials: G
authors.lastname: Rigó
authors.forename: Laura
authors.initials: L
authors.lastname: Zsigmond
authors.forename: Jolán
authors.initials: J
authors.lastname: Csiszár
authors.forename: Ferhan
authors.initials: F
authors.lastname: Ayaydin
authors.forename: Nicolai
authors.initials: N
authors.lastname: Strizhov
authors.forename: Jan
authors.initials: J
authors.lastname: Jásik
authors.forename: Elmon
authors.initials: E
authors.lastname: Schmelzer
authors.forename: Csaba
authors.initials: C
authors.lastname: Koncz
authors.forename: László
authors.initials: L
authors.lastname: Szabados
mesh_terms: D050842:1-Pyrroline-5-Carboxylate Dehydrogenase / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D020131:Genes, Duplicate; D011392:Proline / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D018749:RNA, Plant / Q000187:drug effects / Q000378:metabolism; D014882:Water-Electrolyte Balance
chemical_list: D029681:Arabidopsis Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D011392:Proline; D050842:1-Pyrroline-5-Carboxylate Dehydrogenase