Characterization of regions within the N-terminal 6-kilodalton domain of phytochrome A that modulate its biological activity.
Phytochrome A (phyA) is a red/far-red (FR) light photoreceptor responsible for initiating numerous light-mediated plant growth and developmental responses, especially in FR light-enriched environments. We previously showed that the first 70 amino acids of the polypeptide contain at least two regions with potentially opposite functions (E.T. Jordan, J.R. Cherry, J.M. Walker, R.D. Vierstra [1996] Plant J 9: 243-257). One region is required for activity and correct apoprotein/chromophore interactions, whereas the second appears to regulate phytochrome activity. We have further resolved these functional regions by analysis of N-terminal deletion and alanine-scanning mutants of oat (Avena sativa) phyA in transgenic tobacco (Nicotiana tabacum). The results indicate that the region involved in chromophore/apoprotein interactions contains two separate segments (residues 25-33 and 50-62) also required for biological activity. The region that regulates phyA activity requires only five adjacent serines (Sers) (residues 8-12). Removal or alteration of these Sers generates a photoreceptor that increases the sensitivity of transgenic seedlings to red and FR light more than intact phyA. Taken together, these data identify three distinct regions in the N-terminal domain necessary for photoreceptor activity, and further define the Ser-rich region as an important site for phyA regulation.
Jordan
E T
ET
Department of Horticulture, University of Wisconsin, Madison 53706, USA.
Marita
J M
JM
Clough
R C
RC
Vierstra
R D
RD
D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D018554:Avena / Q000235:genetics; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004307:Dose-Response Relationship, Radiation; D018546:Hypocotyl / Q000528:radiation effects; D008027:Light; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D010834:Phytochrome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051942:Phytochrome A; D030821:Plants, Genetically Modified / Q000528:radiation effects; D010947:Plants, Toxic; D011487:Protein Conformation; D017384:Sequence Deletion; D015398:Signal Transduction; D013053:Spectrophotometry; D014026:Nicotiana / Q000235:genetics / Q000528:radiation effects
D051942:Phytochrome A; D010834:Phytochrome; D000409:Alanine