INVESTIGATORS: E.P. Beers, A.M. Brunner, A.W.Dickerman
INSTITUTION: Virginia Tech
NON-TECHNICAL SUMMARY: Proteins are molecular machines that are requiredfor nearly all biological functions. To function, proteins must interactwith other molecules such as carbohydrates, lipids, nucleic acids andother proteins. Very little is known about the protein-protein interactionsthat plant cells use to control cell wall related biomass production.The identification of protein-protein interaction networks associatedwith biomass production in the woody tissues of poplar, a model biomasscrop, will lead to a more detailed understanding of the molecular biologyand genomics of plant biomass production and ultimately contributeto strategies for biomass crop improvement.
OBJECTIVE: The objective of the proposal is to map protein-proteininteractions relevant to biomass production by focusing on proteinsexpressed in poplarwood, the site of the majority of secondary cell wall synthesis andhence biomass accumulation in poplar.
APPROACH: We have identified approximately 250 poplar genes specificallyassociated with wood formation, and have designated these as thepoplar biomass gene set. Each of these genes will be cloned andexpressed as protein in a yeast expression system known as the yeasttwo-hybridsystemfor protein-protein interaction. In the yeast two-hybrid system,individualproteins can be tested for their ability to interact with otherproteins expressed in the same yeast cell. We will perform a 250 x250 yeasttwo-hybrid matrix assay that will identify interactions occurringbetween any twomembers of the poplar biomass protein set. Selected members ofthe biomass protein set will also be tested in the yeast two-hybridsystemfor theirability to interact with any protein found in a library of proteinsderived from all the genes expressed in poplar wood-forming tissues.Finally,a small number of biomass proteins will be expressed in plantsand tested for their ability to form complexes with other proteinspresentin woodytissue. Members of protein complexes isolated from plants willbe identified using mass spectrometry. All of the interactions revealedas a resultof the three types of protein-protein interaction studies willbecombined to produce a biomass protein-protein interaction map toaid in thecomprehensive understanding of how protein-protein interactionscontribute to biomassproduction by plants.
Name: E.P. Beers