Pr-specific phytochrome phosphorylation in vitro by a protein kinase present in anti-phytochrome maize immunoprecipitates.
Protein kinase activity has repeatedly been found to co-purify with the plant photoreceptor phytochrome, suggesting that light signals received by phytochrome may be transduced or modulated through protein phosphorylation.  In this study immunoprecipitation techniques were used to characterize protein kinase activity associated with phytochrome from maize (Zea mays L.).  A protein kinase that specifically phosphorylated phytochrome was present in washed anti-phytochrome immunoprecipitates of etiolated coleoptile proteins.  No other substrate tested was phosphorylated by this kinase.  Adding salts or detergents to disrupt low-affinity protein interactions reduced background phosphorylation in immunoprecipitates without affecting phytochrome phosphorylation, indicating that the protein kinase catalytic activity is either intrinsic to the phytochrome molecule or associated with it by high-affinity interactions.  Red irradiation (of coleoptiles or extracts) sufficient to approach photoconversion saturation reduced phosphorylation of immunoprecipitated phytochrome.  Subsequent far-red irradiation reversed the red-light effect.  Phytochrome phosphorylation was stimulated about 10-fold by a co-immunoprecipitated factor.  The stimulatory factor was highest in immunoprecipitates when Mg2+ was present in immunoprecipitation reactions but remained in the supernatant in the absence of Mg2+.  These observations provide strong support for the hypothesis that phytochrome-associated protein kinase modulates light responses in vivo.  Since only phytochrome was found to be phosphorylated, the co-immunoprecipitated protein kinase may function to regulate receptor activity.
Biermann
B J
BJ
Department of Plant Biology, University of California, Berkeley 94720, USA.
Pao
L I
LI
Feldman
L J
LJ
D000906:Antibodies / Q000032:analysis; D018548:Cotyledon / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism / Q000528:radiation effects; D008027:Light; D008274:Magnesium; D010766:Phosphorylation; D018462:Photobiology; D010834:Phytochrome / Q000032:analysis / Q000378:metabolism*; D011233:Precipitin Tests; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology* / Q000528:radiation effects; D013053:Spectrophotometry; D003313:Zea mays / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism* / Q000528:radiation effects
D000906:Antibodies; D010834:Phytochrome; D011494:Protein Kinases; D008274:Magnesium