Present address: Dept. of Organismic and Evolutionary Biology, Harvard University, Cambridge MA 02138, USA.
Genome-wide transposon tagging reveals location-dependent effects on transcription and chromatin organization in Arabidopsis
Article first published online: 12 APR 2008
© 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd
The Plant Journal
Volume 55, Issue 3, pages 514–525, August 2008
How to Cite
Rosin, F. M., Watanabe, N., Cacas, J.-L., Kato, N., Arroyo, J. M., Fang, Y., May, B., Vaughn, M., Simorowski, J., Ramu, U., McCombie, R. W., Spector, D. L., Martienssen, R. A. and Lam, E. (2008), Genome-wide transposon tagging reveals location-dependent effects on transcription and chromatin organization in Arabidopsis. The Plant Journal, 55: 514–525. doi: 10.1111/j.1365-313X.2008.03517.x
- Issue published online: 23 JUL 2008
- Article first published online: 12 APR 2008
- Received 2 February 2008; revised 22 March 2008; accepted 26 March 2008; published online 2 June 2008.
Figure S1. Structure of T-DNAs used to create the CCV lines in Arabidopsis is shown. (a) JM71, Ethanol-inducible EYFP-LacI-NLS fusion protein. RB, LB, right and left borders; EYFP-LacI-NLS; enhanced yellow florescent protein-lac repressor-nuclear localization signal; pAlcA,Aspergillus nidulans AlcA promoter; NPTII, neomycin phosphotransferase gene cassette; p35S, CaMV 35S promoter; AlcR, A. nidulans ALCR transcription factor. (b) EL700S, Dex-inducible mGFP (Green fluorescent protein)-LacI-NLS. GVG, glucocorticoid-binding domain-VP16 acidic activation domain-Gal4 DNA-binding domain with pea rbcs-E9 terminator, HPT, hygromycin phosphotransferase with NOS promoter and terminator; 6xUAS, 6xGal4 UAS; TATA, TATA box; with pea rbcs-3A terminator.
Figure S2. RLA in shoots after AZA treatment. Treatment with the DNA methylation inhibitor AZA did not alter the RLA in shoots for CCT lines on the short arm of Ch2. RLA is calculated by setting the observed activity from shoot tissue of our control line, CCP4-20, to 100. Data represents the mean ± SE from triplicate assay samples.
Figure S3. Chromatin dynamics are altered for roots, but not leaves, treated with AZA. Overall mean squared differences in distances < Dd2 > (y-axis, μm2) between LacO spots plotted against the time interval, Dt (x-axis, min) for CCP4.211 root (a), CCP4.211, CCT431, CCT396, and CCT432 leaf (b–e). Lines are black (MS roots), blue (AZA roots), gray (MS leaf) or green (AZA leaf) to indicate treatment and tissue. Data sets represent the mean ± SE of n pairs of distances for each line. Following AZA treatment, CCP4.211 root endoreduplicated nuclei undergo changes in chromatin dynamics with an increased diffusion coefficient and radius of confinement. Treatment with AZA did not greatly affect the confinement radius or diffusion coefficient in leaf nuclei for CCP4.211, CCT431, and CCT396; however, AZA leaves of CCT432 exhibited greater mobility compared to MS leaves.
Table S1. Primers used for locus-specific PCR and ABRC stock numbers are listed for the CCP4, CCP5 and CCT lines.
Table S2. Chromosomal position, relative luciferase activity (RLA) and overall level of GUS activity is given for each line in the collection.
Appendix S1. Protocol: more details of the construction of the CCP4 cassette.
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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.