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Color recovery after photoconversion of H2B::mEosFP allows detection of increased nuclear DNA content in developing plant cells.
Plant Physiol 2012; 158(1):95-106PP

Abstract

Many higher plants are polysomatic whereby different cells possess variable amounts of nuclear DNA. The conditional triggering of endocycles results in higher nuclear DNA content (C value) that in some cases has been correlated to increased cell size. While numerous multicolored fluorescent protein (FP) probes have revealed the general behavior of the nucleus and intranuclear components, direct visualization and estimation of changes in nuclear-DNA content in live cells during their development has not been possible. Recently, monomeric Eos fluorescent protein (mEosFP) has emerged as a useful photoconvertible protein whose color changes irreversibly from a green to a red fluorescent form upon exposure to violet-blue light. The stability and irreversibility of red fluorescent mEosFP suggests that detection of green color recovery would be possible as fresh mEosFP is produced after photoconversion. Thus a ratiometric evaluation of the red and green forms of mEosFP following photoconversion could be used to estimate production of a core histone such as H2B during its concomitant synthesis with DNA in the synthesis phase of the cell cycle. Here we present proof of concept observations on transgenic tobacco (Nicotiana tabacum) Bright Yellow 2 cells and Arabidopsis (Arabidopsis thaliana) plants stably expressing H2B::mEosFP. In Arabidopsis seedlings an increase in green fluorescence is observed specifically in cells known to undergo endoreduplication. The detection of changes in nuclear DNA content by correlating color recovery of H2B::mEosFP after photoconversion is a novel approach involving a single FP. The method has potential for facilitating detailed investigations on conditions that favor increased cell size and the development of polysomaty in plants.

Authors+Show Affiliations

Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G2W1.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22108524

Citation

Wozny, Michael, et al. "Color Recovery After Photoconversion of H2B::mEosFP Allows Detection of Increased Nuclear DNA Content in Developing Plant Cells." Plant Physiology, vol. 158, no. 1, 2012, pp. 95-106.
Wozny M, Schattat MH, Mathur N, et al. Color recovery after photoconversion of H2B::mEosFP allows detection of increased nuclear DNA content in developing plant cells. Plant Physiol. 2012;158(1):95-106.
Wozny, M., Schattat, M. H., Mathur, N., Barton, K., & Mathur, J. (2012). Color recovery after photoconversion of H2B::mEosFP allows detection of increased nuclear DNA content in developing plant cells. Plant Physiology, 158(1), pp. 95-106. doi:10.1104/pp.111.187062.
Wozny M, et al. Color Recovery After Photoconversion of H2B::mEosFP Allows Detection of Increased Nuclear DNA Content in Developing Plant Cells. Plant Physiol. 2012;158(1):95-106. PubMed PMID: 22108524.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Color recovery after photoconversion of H2B::mEosFP allows detection of increased nuclear DNA content in developing plant cells. AU - Wozny,Michael, AU - Schattat,Martin H, AU - Mathur,Neeta, AU - Barton,Kiah, AU - Mathur,Jaideep, Y1 - 2011/11/22/ PY - 2011/11/24/entrez PY - 2011/11/24/pubmed PY - 2012/7/11/medline SP - 95 EP - 106 JF - Plant physiology JO - Plant Physiol. VL - 158 IS - 1 N2 - Many higher plants are polysomatic whereby different cells possess variable amounts of nuclear DNA. The conditional triggering of endocycles results in higher nuclear DNA content (C value) that in some cases has been correlated to increased cell size. While numerous multicolored fluorescent protein (FP) probes have revealed the general behavior of the nucleus and intranuclear components, direct visualization and estimation of changes in nuclear-DNA content in live cells during their development has not been possible. Recently, monomeric Eos fluorescent protein (mEosFP) has emerged as a useful photoconvertible protein whose color changes irreversibly from a green to a red fluorescent form upon exposure to violet-blue light. The stability and irreversibility of red fluorescent mEosFP suggests that detection of green color recovery would be possible as fresh mEosFP is produced after photoconversion. Thus a ratiometric evaluation of the red and green forms of mEosFP following photoconversion could be used to estimate production of a core histone such as H2B during its concomitant synthesis with DNA in the synthesis phase of the cell cycle. Here we present proof of concept observations on transgenic tobacco (Nicotiana tabacum) Bright Yellow 2 cells and Arabidopsis (Arabidopsis thaliana) plants stably expressing H2B::mEosFP. In Arabidopsis seedlings an increase in green fluorescence is observed specifically in cells known to undergo endoreduplication. The detection of changes in nuclear DNA content by correlating color recovery of H2B::mEosFP after photoconversion is a novel approach involving a single FP. The method has potential for facilitating detailed investigations on conditions that favor increased cell size and the development of polysomaty in plants. SN - 1532-2548 UR - https://www.unboundmedicine.com/medline/citation/22108524/Color_recovery_after_photoconversion_of_H2B::mEosFP_allows_detection_of_increased_nuclear_DNA_content_in_developing_plant_cells_ L2 - http://www.plantphysiol.org/cgi/pmidlookup?view=long&pmid=22108524 DB - PRIME DP - Unbound Medicine ER -