INGEBI   02650
INSTITUTO DE INVESTIGACIONES EN INGENIERIA GENETICA Y BIOLOGIA MOLECULAR "DR. HECTOR N TORRES"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
STUDY OF PHOSPHORYLATED RESIDUES OF LEPRK2 IN TOMATO AND THEIR FUNCTION IN POLLEN-PISTIL INTERACTION
Autor/es:
SALEM, T.; WENGIER, D.; MUSCHIETTI J.
Lugar:
Mar del Plata, Buenos Aires, Argentina
Reunión:
Congreso; XLIII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2007
Institución organizadora:
Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB)
Resumen:
LePRK1 and LePRK2 are two pollen-specific receptor kinases fromSolanum lycopersicum, of pollen tubes and potentially involved inpollen-pistil interactions. Transduction, by their kinase activity, of apistil signal might initiate a signalling cascade in the pollen tube thatregulates its growth. LePRK2 is phosphorylated in vivo in pollenmembranes.To determine which LePRK2 residues are phosphorylated in vivo,we separated proteins from germinated pollen, by 2D gels. Tovisualize the phosphorylated isoforms, we performed western blotsusing an antibody against LePRK2. Spots corresponding to thoseisoforms were isolated from a Coomassie blue stained2Dgel.Another strategy involves computational approaches to predictphosphorylation sites in LePRK2.We selected 8 serines or treoninesof LePRK2: S277, S279, S282, S304, S307, T308, T358 and S396.To study their functional relevance during pollen tube growth, wesubstituted them by Alanines using wild-type pLAT52-LePRK2-eGFP vector in mutagenesis, followed by transient expression intobacco pollen. We previously showed pollen tubes that receivedwild type LePRK2-eGFP, displayed aberrant ballon tip morphologyat germination. On the contrary, pollen tubes transfected with theconstruct that had A277, 279, 282, recovered the wild typephenotype. These results will be further studied and complementedby the proteomic approach.