Research Projects with Alfalfa and other plants

Histone Acetylation and Synthesis in Alfalfa and other Higher Plants

(1990-present, University of Missouri-Kansas City)

1. Kapros, T, Kelemen, Z., Waterborg, J.H. and Dudits, D. Short polypyrimidine/purine sequences modulate tissue-specific control of a cell cycle variant alfalfa histone H3 gene promoter Plant Sc. 168: 789-795, 2005.
2. Waterborg, J.H. Dynamics of Histone Acetylation in vivo. A Function for Histone Acetylation? Biochem. Cell Biol. 80: 363-378, 2002.
3. Waterborg, J.H. and Kapros, T. Kinetic Analysis of Histone Acetylation Turnover and Trichostatin A induced Hyper- and Hypoacetylation in Alfalfa Biochem. Cell Biol. 80: 279-293, 2002.
4. Kelemen, Z., Mai, A., Kapros, T., Feher, A., Gyorgyey, J., Waterborg, J.H. and Dudits, D. Transformation vector based on promoter and intron sequences of a replacement histone H3 gene. A tool for high, constitutive gene expression in plants Transgenic Res. 11: 69-72, 2002.
5. Chaubet-Gigot, N., Kapros, T., Flenet, M., Kahn, K., Gigot, C. and Waterborg, J.H. Tissue-dependent enhancement of transgene expression by introns of replacement histone H3 genes of Arabidopsis Plant Mol. Biol. 45: 17-30, 2001.
6. Waterborg, J.H. Histone Synthesis and Turnover in Alfalfa. Fast Loss of Highly Acetylated Replacement Histone H3.2 J. Biol. Chem. 268: 4912-4917, 1993. (Abstract) Slides
7. Waterborg. J.H. Dynamic Methylation of Alfalfa Histone H3 J. Biol. Chem. 268: 4918-4921, 1993. (Abstract)
8. Waterborg, J.H. Sequence Analysis of Acetylation and Methylation of Two Histone H3 Variants of Alfalfa J. Biol. Chem. 265: 17157-17161, 1990. (Abstract)
9. Waterborg, J.H. Multiplicity of Histone H3 Variants in Wheat, Barley, Rice and Maize Plant Physiol. 96: 453-458, 1991.
10. Waterborg, J.H. Existence of Two Histone H3 Variants in Dicotyledonous Plants and Correlation between their Acetylation and Plant Genome Size Plant Mol. Biol. 18: 181-187, 1992.
11. Waterborg, J.H. Histone Variants in the Cereals Rice Biotechnology Quarterly 9: 12-13, 1992.
12. Waterborg, J.H. Identification of Five Sites of Acetylation in Alfalfa Histone H4 Biochemistry 31: 6211-6219, 1992.

Unpublished experiments on dynamic histone acetylation in alfalfa:
Slide show presentation at the 20th West Coast Chromatin and Chromosomes Meeting, Asilomar CA, December 1998:
1. Turnover rates of acetylation of alfalfa core histones
2. Fraction of acetylated forms of alfalfa histones subject to turnover
3. Trichostatin A (TSA) induces hyperacetylation of histone H3, H4 and H2B
4. Histone H3 hyperacetylation by TSA may be limited by irreversible lysine methylation
5. TSA induces transient HYPERacetylation of alfalfa histones followed by HYPOacetylation
   Deacetylase inhibitor Trichostatin A is unstable in alfalfa plant cells !
6. TSA re-addition restores hyperacetylation
7. The balance between acetylating and deacetylating activities is different for H3, H4 and H2B histones in alfalfa chromatin

Cloning and Characterization of Histone H3 Genes in Alfalfa

(1995-1997, University of Missouri-Kansas City)

1. Waterborg, J.H. and Robertson, A.J. Common Features of Analogous Replacement Histone H3 Genes in Animals and Plants J. Mol. Evol. 43: 194-206, 1996. Slides
2. Robertson, A.J., Kapros, T., Dudits, D. and Waterborg, J.H. Identification of the Three Highly Expressed Replacement Histone H3 Genes of Alfalfa DNA Sequence 6: 137-146, 1996.
3. Robertson, A.J., Kapros, T. and Waterborg, J.H. A Cell Cycle-Regulated Histone H3 Gene of Alfalfa with an Atypical Promoter Structure DNA Sequence 7: 209-216, 1997.
4. Kapros, T., Robertson, A.J. and Waterborg, J.H. Histone H3 Transcript Stability in Alfalfa Plant Mol. Biol. 28: 901-914, 1995.

Slide show: Identification of Replacement H3 genes in plants with phylogeny of ALL known H3 genes:
1. Alfalfa has 2 histone H3 variants, H3.1 and H3.2
2. Genes cloned: intron-less H3.1 and intron-bearing H3.2 genes
3. H3.1 genes are expressed in S phase; H3.2 genes are constitutive
4. H3.2 mRNA and protein synthesis is unexpectedly high
5. Newly synthesized histone H3.2 protein is NOT stable
6. 60% of new histone H3.2 protein turns over with a half-life of less than 20 hours (new, unpublished data)
7. Histone H3.1 appears inherently to exist in more stable nucleosomes (new, unpublished data)
8. Most highly acetylated new H3.2 protein is lost fastest
9. In G1 phase-arrested cells: highly acetylated new H3 protein is most labile (new, unpublished data)
   These observations inherently demonstrate that transcription of chromatin in plants causes displacement and loss of nucleosomes at a rate higher than observed in animals !
   See the arguments for this deduction in another slide show.
10. Comparison of intron-less and intron-bearing histone H3 genes in animals and plants
11. Multicellularity in animals and plants originated independently
12. Multicellularity with terminal cell differentiation requires histone protein to maintain chromatin integrity in non-cycling cells
13. Distinct Replacement H3 genes arose, independently, in multicellular animals and plants
14. Amino acid residues 31 and 87 in histone H3 may confer distinct functions to histone H3 variant proteins

Histone Acetylation in Alfalfa

(1984-1988, University of Nevada at Reno)

1. Waterborg, J.H., Harrington, R.E. and Winicov, I. Histone Variants and Acetylated Species from the Alfalfa Plant Medicago sativa Arch. Biochem. Biophys. 256: 167-178, 1987.
2. Winicov, I, Maki, D.H., Waterborg, J.H., Riehm, M.R. and Harrington, R.E. Characterization of the Alfalfa (Medicago sativa) Genome by DNA Reassociation Plant Mol. Biol. 10: 369-371, 1988.
3. Waterborg, J.H., Harrington, R.E. and Winicov, I. Differential Histone Acetylation in Alfalfa (Medicago sativa) Due to Growth in NaCl: Responses in Salt Stressed and Salt Tolerant Callus Cultures Plant Physiol. 90: 237-245, 1989.
4. Winicov, I., Waterborg, J.H., Harrington, R.E. and McCoy, T.J. Messenger RNA Induction and Repression in Cellular Salt Tolerance of Alfalfa (Medicago sativa) Plant Cell Rep. 8: 8-11, 1989.
5. Waterborg, J.H., Harrington, R.E. and Winicov, I. Dynamic Histone Acetylation in Alfalfa Cells. Butyrate Interference With Acetate Labeling Biochim. Biophys. Acta 1049: 324-330, 1990.