Mutational analysis of phytochrome B identifies a small COOH-terminal-domain region critical for regulatory activity.
Overexpression of phytochrome B (phyB) in transgenic Arabidopsis results in enhanced deetiolation in red light. To define domains of phyB functionally important for its regulatory activity, we performed chemical mutagenesis of a phyB-overexpressing line and screened for phenotypic revertants in red light. Four phyB-transgene-linked revertants that retain parental levels of full-length, dimeric, and spectrally normal overexpressed phyB were identified among 101 red-light-specific revertants. All carry single amino acid substitutions in the transgene-encoded phyB that reduce activity by 40- to 1000-fold compared to the nonmutagenized parent. The data indicate that the mutant molecules are fully active in photosignal perception but defective in the regulatory activity responsible for signal transfer to downstream components. All four mutations fall within a 62-residue region in the COOH-terminal domain of phyB, with two independent mutations occurring in a single amino acid, Gly-767. Accumulating evidence indicates that the identified region is a critical determinant in the regulatory function of both phyB and phyA.
Wagner
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Department of Plant Biology, University of California, Berkeley 94720, USA.
Quail
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D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000528:radiation effects; D029681:Arabidopsis Proteins; D004252:DNA Mutational Analysis; D008027:Light; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010786:Photoreceptor Cells; D010834:Phytochrome / Q000235:genetics* / Q000528:radiation effects; D051939:Phytochrome B; D030821:Plants, Genetically Modified; D012641:Selection, Genetic; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000235:genetics*; D013329:Structure-Activity Relationship; D014157:Transcription Factors
D029681:Arabidopsis Proteins; C497952:PHYB protein, Arabidopsis; D014157:Transcription Factors; D010834:Phytochrome; D051939:Phytochrome B