Organization, expression and nucleotide sequence of the operon encoding R-phycoerythrin alpha and beta subunits from the red alga Polysiphonia boldii.
The characterization of the operon encoding the alpha and beta subunits of rhodophytan (R)-phycoerythrin (PE) from the macrophytic red alga Polysiphonia boldii is reported. This plastid-encoded operon was cloned, its nucleotide sequence determined, and its expression characterized by northern and primer extension analyses. The arrangement and expression of the PE alpha and beta genes, named rpeA and rpeB, are similar to those of the cyanobacterial (C)-PE genes: rpeB is located 5' of rpeA, with an intergenic region of 64 nucleotides. The two genes are transcribed on a 1.25 kb dicistronic transcript, and each coding region is preceded by a prokaryotic ribosome binding site consensus sequence. Transcription is initiated 95 nucleotides upstream of the initiating methionine codon of rpeB. The promoter region resembles that of prokaryotic genes, with an AT-rich -10 sequence. A direct pentanucleotide repeat (5'-TGTTA-3') was found in the -35 region. This pentanucleotide is present upstream of all PE operons that have been characterized thus far. An extensive inverted repeat is present 3' of rpeA; inverted repeats are found downstream of all PE operons sequenced to date, although the sequence is not conserved. The deduced amino acid sequences from these genes provide complete sequences for an R-PE. Of the amino acid residues 85% are identical to those of bangeophycean (B)-PE from the unicellular red alga Porphyridium cruentum. Conserved residues include cysteines at the bilin attachment sites of C- and B-PEs, aspartates at positions postulated to interact with bilin chromophores, and an apparent consensus sequence for N-methylation of an asparagine residue in C-PEs.
Roell
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Marine Biotechnology Center, University of California, Santa Barbara 93106.
Morse
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017343:Genes, Plant; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009876:Operon; D010799:Phycoerythrin / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D000461:Rhodophyta / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D010799:Phycoerythrin; D004247:DNA