DamID, a new tool for studying plant chromatin profiling in vivo, and its use to identify putative LHP1 target loci
Article first published online: 30 AUG 2006
The Plant Journal
Volume 48, Issue 1, pages 153–163, October 2006
How to Cite
Germann, S., Juul-Jensen, T., Letarnec, B. and Gaudin, V. (2006), DamID, a new tool for studying plant chromatin profiling in vivo, and its use to identify putative LHP1 target loci. The Plant Journal, 48: 153–163. doi: 10.1111/j.1365-313X.2006.02859.x
- Issue published online: 30 AUG 2006
- Article first published online: 30 AUG 2006
- Received 26 April 2006; revised 12 June 2006; accepted 27 June 2006.
- DNA–protein interaction
We show here that the in vivo methylation-based tagging technique DamID (DNA adenine methyltransferase identification) can be used for studies of DNA–protein interactions or chromatin profiling in plants. We have demonstrated the feasibility, reproducibility and sensitivity of the method in Arabidopsis thaliana, using the well-known yeast GAL4 transcription factor, for which DNA-binding sites (UASG) were introduced into the plant genome. We monitored the methylation resulting from the activity of DNA adenine methyltransferase fused to the protein of interest, by combining digestion with methylation-sensitive restriction enzymes and quantitative PCR. We then used DamID to identify genomic targets of LHP1, a protein mostly associated with euchromatin. We showed that LHP1 was targeted to the promoter and transcribed regions of four genes: AG, AP3, FT and PI. Our data also demonstrate that LHP1, like its animal homologues, has a high binding affinity for A/T-rich regions, binding particularly strongly to the large regulatory introns of AG and PI. We identified three major characteristics of LHP1 binding, highlighting the similarities between plant LHP1 and animal HP1 proteins.