Effect of overexpression of Arabidopsis damaged DNA-binding protein 1A on de-etiolated 1

Planta. 2010 Jan;231(2):337-48. doi: 10.1007/s00425-009-1056-6. Epub 2009 Nov 18.

Abstract

In Arabidopsis thaliana, de-etiolated 1 mutants (det1) grown in the dark resemble light-grown wild-type seedlings. Arabidopsis DET1 encodes a 62 kD protein, which is a negative regulator of light signaling. UV-damaged DNA-binding protein 1 (DDB1) was initially identified due to its role in human DNA damage repair. Arabidopsis has two DDB1 homologs: DDB1A and DDB1B. DDB1A mutation enhances det1 mutant phenotypes. In this study, we generated Arabidopsis lines that overexpress DDB1A-3HA in wild-type, det1, as well as Myc-DET1 or GFP-DET1 rescued genetic backgrounds. DDB1A-3HA overexpression resulted in decreased apical hook formation in wild-type dark-grown seedlings, and enhanced det1 small rosette and early flowering time phenotypes. In the Myc-DET1 background, DDB1A-3HA overexpression resulted in decreased rescue of dark- and light-grown hypocotyl length, light-grown anthocyanin and chlorophyll levels, adult height and stem number phenotypes. This result is consistent with the decreased levels of Myc-DET1 protein detected in the DDB1A-3HA overexpression line. The GFP-DET1 DDB1A-3HA double overexpression line exhibited increased rescue of dark and light-grown hypocotyl length and light-grown chlorophyll level phenotypes relative to GFP-DET1 alone, despite the fact that GFP-DET1 protein also decreased in the double overexpression line. In addition, increased DET1 resulted in decreased DDB1A-3HA levels due to proteasomal degradation. Overall, DDB1A-3HA overexpression affected phenotypes in a variety of DET1 backgrounds, reduced epitope-tagged DET1 levels, and, correlatively, in general dampened the rescue of det1 mutants by the DET1-DDB1A complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / drug effects
  • Arabidopsis / genetics
  • Arabidopsis / growth & development
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Chromatography, Gel
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Darkness
  • Gene Expression Regulation, Plant / drug effects
  • Green Fluorescent Proteins / metabolism
  • Intracellular Signaling Peptides and Proteins
  • Leupeptins / pharmacology
  • Nuclear Proteins / metabolism*
  • Phenotype
  • Proteasome Inhibitors
  • Protein Binding / drug effects
  • Protein Transport / drug effects
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Seedlings / drug effects
  • Seedlings / genetics
  • Seedlings / growth & development

Substances

  • Arabidopsis Proteins
  • DDB1a protein, Arabidopsis
  • DET1 protein, Arabidopsis
  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Leupeptins
  • Nuclear Proteins
  • Proteasome Inhibitors
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde