Identification and characterization of transcription factors regulating Arabidopsis HAK5

Plant Cell Physiol. 2013 Sep;54(9):1478-90. doi: 10.1093/pcp/pct094. Epub 2013 Jul 2.

Abstract

Potassium (K) is an essential macronutrient for plant growth and reproduction. HAK5, an Arabidopsis high-affinity K transporter gene, plays an important role in K uptake. Its expression is up-regulated in response to K deprivation and is rapidly down-regulated when sufficient K levels have been re-established. To identify transcription factors regulating HAK5, an Arabidopsis TF FOX (Transcription Factor Full-length cDNA Over-eXpressor) library containing approximately 800 transcription factors was used to transform lines previously transformed with a luciferase reporter gene whose expression was driven by the HAK5 promoter. When grown under sufficient K levels, 87 lines with high luciferase activity were identified, and endogenous HAK5 expression was confirmed in 27 lines. Four lines overexpressing DDF2 (Dwarf and Delayed Flowering 2), JLO (Jagged Lateral Organs), TFII_A (Transcription initiation Factor II_A gamma chain) and bHLH121 (basic Helix-Loop-Helix 121) were chosen for further characterization by luciferase activity, endogenous HAK5 level and root growth in K-deficient conditions. Further analysis showed that the expression of these transcription factors increased in response to low K and salt stress. In comparison with controls, root growth under low K conditions was better in each of these four TF FOX lines. Activation of HAK5 expression by these four transcription factors required at least 310 bp of upstream sequence of the HAK5 promoter. These results indicate that at least these four transcription factors can bind to the HAK5 promoter in response to K limitation and activate HAK5 expression, thus allowing plants to adapt to nutrient stress.

Keywords: Arabidopsis TFs FOX; HAK5; Potassium-deficient signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Gene Expression Regulation, Plant / genetics
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Plants, Genetically Modified
  • Potassium / pharmacology
  • Potassium-Hydrogen Antiporters
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sodium Chloride / pharmacology
  • Symporters / genetics*
  • Symporters / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription Factors, TFII / genetics
  • Transcription Factors, TFII / metabolism

Substances

  • Arabidopsis Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • DDF2 protein, Arabidopsis
  • JAGGED LATERAL ORGANS protein, Arabidopsis
  • Potassium-Hydrogen Antiporters
  • Repressor Proteins
  • Symporters
  • Transcription Factors
  • Transcription Factors, TFII
  • bHLH121 protein, Arabidopsis
  • potassium transporter, Arabidopsis
  • transcription initiation factor IIA subunit 2, Arabidopsis
  • Sodium Chloride
  • Luciferases
  • Glucuronidase
  • Potassium