Present Address: Monsanto-Calgene Campus, 1920 5th Street, Davis, CA 95616, USA
ORTH/VIM proteins that regulate DNA methylation are functional ubiquitin E3 ligases
Article first published online: 15 AUG 2008
© 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd
The Plant Journal
Volume 56, Issue 5, pages 704–715, December 2008
How to Cite
Kraft, E., Bostick, M., Jacobsen, S. E. and Callis, J. (2008), ORTH/VIM proteins that regulate DNA methylation are functional ubiquitin E3 ligases. The Plant Journal, 56: 704–715. doi: 10.1111/j.1365-313X.2008.03631.x
- Issue published online: 27 NOV 2008
- Article first published online: 15 AUG 2008
- Received 22 April 2008; revised 25 June 2008; accepted 30 June 2008; published online 15 August 2008.
Figure S1. Clustal X generated protein alignment of ORTH proteins and SRA domain. (a) Alignment of the entire protein coding region of the Arabidopsis ORTH proteins. Domains are indicated as follows: blue bar shows position of PHD domain, red bar indicates position of N-terminal RING domain, green bar indicates position of the SRA domain, purple bar indicates position of the C-terminal RING domain. All domain limits are based on SMART predictions. (b) The core SRA domain alignment of all Arabidopsis SRA domain-containing proteins as well as representative orthologues of the ORTH proteins in rice (OsXP471608; OsXP493873), human (ICBP90; NIRF), mouse (NIRF; NP95), Danio (DrNP95like), and Xenopus (XlNP95like) (See Figure 1 for NCBI locus codes for each protein.). SRA domain-only and SRA-SET domain-containing proteins are listed by their AGI code. Arrows indicate positions in ORTH1 and other SRA domain-containing proteins known to disrupt methylated DNA association (Johnson et al., 2007).
Figure S2. Overexpression of GFP-ORL does not cause a late-flowering phenotype. Eight independent insertional lines expressing GFP-ORL under the 35S promoter were analyzed for a flowering time phenotype. The number of leaves produced before the transition to reproductive tissue was counted from at six to twenty segregating T2 individuals. Each bar represents mean ± SEM.
Figure S3. Overexpression of ORTH1 causes a late flowering phenotype. (a) The flowering time of seven to nineteen T2 plants from ten independent 35S::ORTH1 lines and wild type (Col) was measured by counting leaves produced before the transition to reproductive tissue. Each bar represents mean ± SD. Asterisk is above lines that are statistically different from Col based on a student’s t test (P < 0.005). (b) Southern blots for FWA (upper) and Cen180 bp repeats (lower) with DNA pooled from all T2 plants from ten ORTH1 independent plants. The upper band from Col (C) DNA is fully methylated (m), while lower bands are products from unmethylated (u) DNA.
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