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Stony Brook University, State University of New York

5 ARTICLES PUBLISHED IN JoVE

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Biology

A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment
Shoko Ueki 1, Benjamin L. Meyers 1, Farzana Yasmin 2, Vitaly Citovsky 2
1Department of Biochemistry and Cell Biology, State University of New York at Stony Brook, 2Bio-Medical Engineering Department, NED University of Engineering and Technology

Macromolecular trafficking between plant cells can be assessed by transiently expressing a fluorescently-tagged protein of interest and analyzing its intra- and intercellular distribution by confocal microscopy.

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Biology

Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
Shoko Ueki 1, Benoît Lacroix 1, Vitaly Citovsky 1
1Department of Biochemistry and Cell Biology, State University of New York

Testing protein-protein interaction is indispensable for dissection of protein functionality. Here, we introduce an in vitro protein-protein binding assay to probe a membrane-immobilized protein with a soluble protein. This assay provides a reliable method to test interaction between an insoluble protein and a protein in solution.

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Biology

Assaying Proteasomal Degradation in a Cell-free System in Plants
Elena García-Cano 1, Adi Zaltsman 1, Vitaly Citovsky 1
1Department of Biochemistry and Cell Biology, Stony Brook University, State University of New York

Targeted protein degradation represents a major regulatory mechanism for cell function. It occurs via a conserved ubiquitin-proteasome pathway, which attaches polyubiquitin chains to the target protein that then serve as molecular “tags” for the 26S proteasome. Here, we describe a simple and reliable cell-free assay for proteasomal degradation of proteins.

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Immunology and Infection

Identification of Plasmodesmal Localization Sequences in Proteins In Planta
Cheng Yuan 1, Sondra G. Lazarowitz 2, Vitaly Citovsky 1
1Department of Biochemistry and Cell Biology, State University of New York, 2Department of Plant Pathology and Plant-Microbe Biology, Cornell University

Plant intercellular connections, the plasmodesmata (Pd), play central roles in plant physiology and plant-virus interactions. Critical to Pd transport are sorting signals that direct proteins to Pd. However, our knowledge about these sequences is still in its infancy. We describe a strategy to identify Pd localization signals in Pd-targeted proteins.

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Biology

Investigating Interactions Between Histone Modifying Enzymes and Transcription Factors in vivo by Fluorescence Resonance Energy Transfer
Mi Sa Vo Phan 1, Phu Tri Tran 1, Vitaly Citovsky 1
1Department of Biochemistry and Cell Biology, State University of New York

Fluorescence resonance energy transfer (FRET) is an imaging technique for detecting protein interactions in living cells. Here, a FRET protocol is presented to study the association of histone-modifying enzymes with transcription factors that recruit them to the target promoters for epigenetic regulation of gene expression in plant tissues.

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