Cyclin D polynucleotides polypeptides and uses thereof

Abstract

The invention provides isolated polynucleotides and their encoded proteins that are involved in cell cycle regulation. The invention further provides recombinant expression cassettes, host cells, transgenic plants, and antibody compositions. The present invention provides methods and compositions relating to altering cell cycle protein content and/or composition of plants.

What is claimed is  
     
         1 . A method for increasing transformation frequency in a plant cell comprising introducing into the plant cell a Cyclin D (CycD) polypeptide having transformation frequency-increasing activity.  
     
     
         2 . A method for increasing transformation frequency comprising introducing a polynucleotide of interest into a plant cell containing a CycD polypeptide having transformation frequency-increasing activity.  
     
     
         3 . A method for increasing transformation frequency of a plant cell comprising introducing into a plant cell an isolated CycD polynucleotide wherein said CycD polynucleotide is operably linked to a promoter capable of driving expression in the plant cell and wherein a CycD polypeptide is expressed from said CycD polynucleotide, said CycD polypeptide having transformation frequency-increasing activity.  
     
     
         4 . A method for increasing transformation frequency of a plant cell comprising introducing into a plant cell an isolated CycD polynucleotide wherein said CycD polynucleotide is operably linked to a promoter capable of driving expression in said plant cell and wherein a CycD polypeptide is expressed from said CycD polynucleotide, said CycD polypeptide comprising cell cycle modulating activity.  
     
     
         5 . The method of  claim 4  further comprising introducing into the plant cell at least one polynucleotide of interest, said polynucleotide optionally being operably linked to a promoter capable of driving expression in said plant cell.  
     
     
         6 . The method of  claim 4  wherein said CycD polynucleotide encodes a plant CycD polypeptide.  
     
     
         7 . The method of  claim 6  wherein said plant CycD polypeptide is a monocot CycD polypeptide.  
     
     
         8 . The method of  claim 7  wherein said monocot CycD is a maize CycD polypeptide.  
     
     
         9 . The method of  claim 8  wherein said maize CycD is a CycDa or CycDc polypeptide.  
     
     
         10 . The method of  claim 9 , wherein said CycD polypeptide has 90% sequence identity to SEQ ID NOs 2 or 14.  
     
     
         11 . The method of  claim 10 , further comprising a plant CycD specific binding motif, LXCXE and a conserved tryptophan residue in a conserved motif, WILKV.  
     
     
         12 . The method of  claim 4  wherein said CycD polynucleotide has 85% sequence identity to Seq ID Nos 1 or 13.  
     
     
         13 . The Cyc D polynucleotide of  claim 4  wherein the promoter is a constitutive promoter.  
     
     
         14 . The CycD polynucleotide of  claim 4  wherein the promoter is inducible, tissue specific, developmentally regulated, or temporally regulated.  
     
     
         15 . The method of  claim 4  wherein said CycD polynucleotide operably linked to a promoter is comprised in a recombinant expression cassette.  
     
     
         16 . The method of  claim 4 , wherein the plant cell is stably transformed.  
     
     
         17 . The method of  claim 4 , wherein the plant cell is a monocot or a dicot cell.  
     
     
         18 . The method of  claim 17 , wherein said monocot or dicot cell is from corn, soybean, wheat, rice, alfalfa, sunflower, or canola.  
     
     
         19 . The method of clam  16 , further comprising growing the plant cell into a plant.  
     
     
         20 . The method of  claim 4 , wherein increasing the level of CycD polypeptide expression increases the number of transgenic events.  
     
     
         21 . A method of increasing transformation frequency in a plant cell comprising introducing into the plant cell a CycD RNA or polypeptide, wherein said polypeptide comprises cell cycle modulating activity.  
     
     
         22 . A method of modulating the cell cycle comprising transforming the plant cell with a recombinant expression cassette comprising an isolated CycD polynucleotide operably linked to a promoter, wherein said CycD polynucleotide encodes a CycD polypeptide which modulates cell cycle activity.  
     
     
         23 . A method for providing a means of positive selection comprising expressing a CycD polypeptide in a plant cell and selecting for cells exhibiting a positive growth advantage, said polypeptide having transformation frequency increasing activity.  
     
     
         24 . The method of  claim 23  further comprising a polynucleotide of interest.  
     
     
         25 . The method of  claim 1 , wherein the cell comprises said CycD polypeptide from introduced RNA or protein or protein delivered via Agrobacterium.  
     
     
         26 . A method of modulating the level of CycD protein in a cell, comprising: 
 (a) transforming a cell with a recombinant expression cassette comprising a CycD polynucleotide operably linked to a promoter;    (b) growing the cell under cell-growing conditions for a time sufficient to induce expression of the polynucleotide sufficient to modulate CycD protein in the cell.    
     
     
         27 . The method of  claim 26 , wherein CycD protein is increased.  
     
     
         28 . The method of  claim 26 , wherein CycD protein is decreased.  
     
     
         29 . The method of  claim 26 , wherein the level of CycD protein in the cell is transiently modulated by introducing CycD ribonucleic acid.  
     
     
         30 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient to alter cell division.  
     
     
         31 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient to increase the number of dividing cells.  
     
     
         32 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient improves transformation frequencies.  
     
     
         33 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient to alter cell growth.  
     
     
         34 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient to provide a positive growth advantage for the cell.  
     
     
         35 . The method of  claim 26 , wherein the CycD protein is present in an amount sufficient to increase the growth rate.  
     
     
         36 . The method of  claim 26 , wherein the cell is a plant cell and the plant cell is grown under conditions appropriate for regenerating a plant capable of expressing CycD protein.  
     
     
         37 . The method of  claim 36 , wherein the plant cell is from corn, soybean, wheat, rice, alfalfa, sunflower, or canola.  
     
     
         38 . The method of  claim 36 , wherein the CycD protein is present in an amount sufficient to increase crop yield.  
     
     
         39 . The method of  claim 36 , wherein the CycD protein is present in an amount sufficient to alter plant height or size.  
     
     
         40 . The method of  claim 36 , wherein the CycD protein is present in an amount sufficient to enhance or inhibit organ growth.  
     
     
         41 . The method of  claim 39 , wherein the organ is a seed, root, shoot, ear, tassel, stalk, pollen, or stamen.  
     
     
         42 . The method of  claim 40 , wherein the level of CycD protein is modulated to produce organ ablation.  
     
     
         43 . The method of  claim 36 , wherein the level of CycD protein is modulated to produce parthenocarpic fruits.  
     
     
         44 . The method of  claim 36 , wherein the level of CycD protein is modulated to produce male sterile plants.  
     
     
         45 . The method of  claim 42 , wherein the CycD protein is present in an amount sufficient to enhance embryogenic response.  
     
     
         46 . The method of  claim 36 , wherein the CycD protein is present in an amount sufficient to increase callus induction.  
     
     
         47 . The method of  claim 36 , wherein the level of CycD protein is modulated to provide for positive selection.  
     
     
         48 . The method of  claim 36 , wherein the level of CycD protein is modulated to increase plant regeneration.  
     
     
         49 . The method of  claim 42 , wherein the level of CycD protein is modulated to alter the percent of time that the cells are arrested in G1 or G0 phase.  
     
     
         50 . The method of  claim 36 , wherein the level of CycD protein is modulated to alter the amount of time the cell spends in a particular cell cycle.  
     
     
         51 . The method of  claim 43 , wherein the level of CycD protein is modulated to improve the response of the cells to environmental stress including dehydration, heat, or cold.  
     
     
         52 . The method of  claim 36 , wherein the level of CycD protein is modulated to increase the number of pods per plant.  
     
     
         53 . The method of  claim 36 , wherein the level of CycD protein is modulated to increase the number of seeds per pod or ear.  
     
     
         54 . The method of  claim 36 , wherein the level of CycD protein is modulated to alter the lag time in seed development.  
     
     
         55 . The method of  claim 36 , wherein the level of CycD protein is modulated to provide hormone independent cell growth.  
     
     
         56 . The method of  claim 26 , wherein the level of CycD protein is modulated to increase the growth rate of cells in bioreactors.  
     
     
         57 . The method of  claim 26 , wherein the level of CycD protein in cells is: transiently modulated by introducing CycD ribonucleic acid.  
     
     
         58 . A method for transiently modulating the level of CycD protein in plant cells comprising introducing CycD polypeptides.