Reduced function of the RNA-binding protein FPA rescues a T-DNA insertion mutant in the Arabidopsis ZHOUPI gene by promoting transcriptional read-through

Plant Mol Biol. 2016 Jul;91(4-5):549-61. doi: 10.1007/s11103-016-0487-2. Epub 2016 May 10.

Abstract

T-DNA insertion mutants have been widely used to investigate plant gene functions. Unexpectedly, in several reported cases, the phenotype of T-DNA insertion mutations can be suppressed because of trans T-DNA interactions associated with epigenetic modification, which indicates that caution is needed when T-DNA mutants are used. In the present study, we characterized a novel process suppressing a T-DNA mutation. The spz2 (suppressor of zou 2) mutant was isolated as a suppressor of the phenotype of the zou-4 mutant caused by a T-DNA insertion in the first intron. The spz2 mutation partially recovered the native ZOU gene expression in the zou-4 background, but not in two other zou alleles, zou-2 and zou-3, with T-DNAs inserted in the exon and intron, respectively. The suppressed phenotype was inherited in a Mendelian fashion and is not associated with epigenetic modification. The recovery of the native ZOU gene expression in the spz2 zou-4 double mutant is caused by transcriptional read-through of the intronic T-DNA as a result of decreased proximal polyadenylation. SPZ2 encodes an RNA-binding protein, FPA, which is known to regulate polyadenylation site selection. This is the first example of FPA rescuing a T-DNA insertion mutation by affecting the polyadenylation site selection.

Keywords: Alternative polyadenylation; Arabidopsis; FPA; T-DNA suppression; ZOU; spz2.

MeSH terms

  • Alleles
  • Arabidopsis / genetics*
  • Arabidopsis / growth & development
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism*
  • Base Sequence
  • Basic Helix-Loop-Helix Transcription Factors / genetics*
  • Cloning, Molecular
  • DNA, Bacterial / genetics*
  • Drug Resistance, Microbial / genetics
  • Epigenesis, Genetic
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant
  • Genes, Plant*
  • Genes, Suppressor
  • Introns / genetics
  • Mutagenesis, Insertional / genetics*
  • Mutation
  • Phenotype
  • Polyadenylation / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism*
  • Seeds / genetics
  • Seeds / growth & development
  • Transcription, Genetic*

Substances

  • Arabidopsis Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • DNA, Bacterial
  • FPA protein, Arabidopsis
  • RETARDED GROWTH OF EMBRYO1 protein, Arabidopsis
  • RNA, Messenger
  • RNA-Binding Proteins
  • T-DNA