, Volume 236, Issue 1, pp 1–17 | Cite as

A novel HSI2 mutation in Arabidopsis affects the PHD-like domain and leads to derepression of seed-specific gene expression

  • Vijaykumar Veerappan
  • Jing Wang
  • Miyoung Kang
  • Joohyun Lee
  • Yuhong Tang
  • Ajay K. Jha
  • Huazhong Shi
  • Ravishankar Palanivelu
  • Randy D. Allen
Original Article


Two related B3 domain transcriptional repressors, HSI2 (HIGH-LEVEL EXPRESSION OF SUGAR-INDUCIBLE GENE2)/VAL1 (VP1/ABI3-LIKE1) and HSL1 (HSI2-LIKE1)/VAL2, function redundantly to repress key transcriptional regulators of seed maturation genes in Arabidopsis thaliana seedlings. Using a forward genetic screen designed to isolate trans-acting mutants that affected expression of a transgene containing the glutathione S-transferase F8 promoter::luciferase (GSTF8::LUC) reporter, we identified a novel HSI2 mutant allele, hsi2-4, that exhibits constitutively elevated luciferase expression while expression of the endogenous GSTF8 transcript remains unchanged. The hsi2-4 lesion was found to be a missense mutation that results in the substitution of a conserved cysteine within the plant homeodomain-like (PHD) motif of HSI2. Microarray analysis of hsi2-4 and hsi2-4 hsl1 mutants indicated that the HSI2 PHD-like domain functions non-redundantly to repress a subset of seed maturation genes, including those that encode AGL15 (AGAMOUS-LIKE15), FUSCA3 (FUS3), cruciferins, cupin family proteins, late-embryogenesis abundant protein, oleosins, 2S albumins and other seed-specific proteins in Arabidopsis seedlings. Many genes that are responsive to this mutation in the HSI2 PHD-like domain are enriched in histone H3 trimethylation on lysine 27 residues (H3K27me3), a repressive epigenetic mark. Chromatin immunoprecipitation analysis showed that sequences of the GSTF8::LUC transgene are enriched in H3K27me3 in a HSI2 PHD domain-dependent manner. These results indicate that the transcriptional repression activity of the HSI2 PHD domain could be mediated, at least in part, by its participation in the deposition of H3K27me3 on the chromatin of specific target genes.


HSI2 VAL1 PHD-like domain H3K27me3 Seed maturation Transgene silencing 







Plant homeodomain


Wild-type luciferase




Neomycin phosphotransferase II



The authors would like to thank Drs. Uppalapati Rao and Mohamed Fokar for comments on the manuscript. No conflict of interest is declared. This work was supported by a grant from the Samuel Roberts Noble Foundation and an endowment from the Walter Sitlington Foundation to RDA.

Supplementary material

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Supplementary material 4 (XLS 17159 kb)
425_2012_1630_MOESM5_ESM.xls (105 kb)
Supplementary material 5 (XLS 105 kb)


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Vijaykumar Veerappan
    • 1
  • Jing Wang
    • 2
  • Miyoung Kang
    • 1
  • Joohyun Lee
    • 1
  • Yuhong Tang
    • 3
  • Ajay K. Jha
    • 4
  • Huazhong Shi
    • 5
  • Ravishankar Palanivelu
    • 6
  • Randy D. Allen
    • 1
  1. 1.Institute for Agricultural BiosciencesOklahoma State UniversityArdmoreUSA
  2. 2.United States Department of AgricultureCropping Systems Research LaboratoryLubbockUSA
  3. 3.Plant Biology DivisionSamuel Roberts Noble FoundationArdmoreUSA
  4. 4.Department of BotanyOklahoma State UniversityStillwaterUSA
  5. 5.Department of Chemistry and BiochemistryTexas Tech UniversityLubbockUSA
  6. 6.School of Plant SciencesUniversity of ArizonaTucsonUSA

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