The nuclear localization signal is required for the function of squamosa promoter binding protein-like gene 9 to promote vegetative phase change in Arabidopsis

Plant Mol Biol. 2019 Aug;100(6):571-578. doi: 10.1007/s11103-019-00863-5. Epub 2019 Apr 5.

Abstract

A mutation in the nuclear localization signal of squamosa promoter binding like-protein 9 (SPL9) delays vegetative phase change by disrupting its nuclear localization. The juvenile-to-adult phase transition is a critical developmental process in plant development, and it is regulated by a decrease in miR156/157 and a corresponding increase in their targets, squamosa promoter binding protein-like (SPL) genes. SPL proteins contain a conserved SBP domain with putative nuclear localization signals (NLSs) at their C-terminals. Some SPLs promote vegetative phase change by promoting miR172 expression, but the function of nuclear localization signals in those SPLs remains unknown. Here, we identified a loss-of-function mutant, which we named del6, with delayed vegetative phase change phenotypes in a forward genetic screen. Map-based cloning, the whole genome resequencing, and allelic complementation test demonstrate that a G-to-A substitution in the SPL9 gene is responsible for the delayed vegetative phase change phenotypes. In del6, the mutation causes a substitution of the glutamine (Gln) for the conserved basic amino acid arginine (Arg) in the NLS of the SBP domain, and disrupts the normal nuclear localization and function of SPL9. Therefore, our work demonstrates that the NLSs in the SBP domain of SPL9 are indispensable for its nuclear localization and normal function in Arabidopsis.

Keywords: Nuclear localization signals (NLS); SBP domain; SPL9; Vegetative phase change.

MeSH terms

  • Alleles
  • Arabidopsis / genetics*
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / physiology*
  • Arginine / chemistry
  • Carrier Proteins / metabolism
  • Cell Nucleus / metabolism
  • Cloning, Molecular
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Plant
  • Genetic Vectors
  • Genome, Plant
  • Glutamine / chemistry
  • MicroRNAs / genetics
  • Mutation
  • Nuclear Localization Signals*
  • Phenotype
  • Plant Leaves / metabolism
  • Plants, Genetically Modified
  • Promoter Regions, Genetic*
  • Trans-Activators / genetics*
  • Trans-Activators / physiology*

Substances

  • Arabidopsis Proteins
  • Carrier Proteins
  • MIRN157 microRNA, Arabidopsis
  • MicroRNAs
  • Mirn156 microRNA, Arabidopsis
  • Nuclear Localization Signals
  • SPL9 protein, Arabidopsis
  • Trans-Activators
  • Glutamine
  • Arginine