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Plant Cell, Vol. 13, 425-436, February 2001, Copyright © 2001, American Society of Plant Physiologists

The Arabidopsis HY2 Gene Encodes Phytochromobilin Synthase, a Ferredoxin-Dependent Biliverdin Reductase

Takayuki Kohchia, Keiko Mukougawaa, Nicole Frankenbergb, Munehisa Masudaa, Akiho Yokotaa, and J. Clark Lagariasb
a Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan
b Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, California 95616

Correspondence to: Takayuki Kohchi, kouchi{at}bs.aist-nara.ac.jp (E-mail), 81-743-72-5569 (fax)

Light perception by the plant photoreceptor phytochrome requires the tetrapyrrole chromophore phytochromobilin (P{Phi}B), which is covalently attached to a large apoprotein. Arabidopsis mutants hy1 and hy2, which are defective in P{Phi}B biosynthesis, display altered responses to light due to a deficiency in photoactive phytochrome. Here, we describe the isolation of the HY2 gene by map-based cloning. hy2 mutant alleles possess alterations within this locus, some of which affect the expression of the HY2 transcript. HY2 encodes a soluble protein precursor of 38 kD with a putative N-terminal plastid transit peptide. The HY2 transit peptide is sufficient to localize the reporter green fluorescent protein to plastids. Purified mature recombinant HY2 protein exhibits P{Phi}B synthase activity (i.e., ferredoxin-dependent reduction of biliverdin IX{alpha} to P{Phi}B), as confirmed by HPLC and by the ability of the bilin reaction products to combine with apophytochrome to yield photoactive holophytochrome. Database searches and hybridization studies suggest that HY2 is a unique gene in the Arabidopsis genome that is related to a family of proteins found in oxygenic photosynthetic bacteria.


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