A 2.4 kilobase-pair segment of the spinach plastid chromosome carrying the genes for apocytochrome b6 and subunit 4 of the thylakoid membrane cytochrome b/f complex has been analysed by DNA sequencing and Northern blot analysis. The nucleotide sequence reveals two uninterrupted open reading frames o
Nucleotide sequence of the clustered genes for the 44 kd chlorophyllaapoprotein and the “32 kd”-like protein of the photosystem II reaction center in the spinach plastid chromosome
✍ Scribed by Juliane Alt; Julia Morris; Peter Westhoff; Reinhold G. Herrmann
- Publisher
- Springer-Verlag
- Year
- 1984
- Tongue
- English
- Weight
- 1009 KB
- Volume
- 8
- Category
- Article
- ISSN
- 0172-8083
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✦ Synopsis
A 2,900 base pair DNA segment of the spinach plastid chromosome which encodes the genes for the 44 kd chlorophyll a apoprotein and a "32 kd"-like protein of the photosystem II reaction center has been subjected to sequence and Northern blot analysis. The genes are located almost centrally in the large single-copy segment of the chromosome adjacent to the two genes for the P700 chlorophyll a apoproteins of the photosystem I reaction center. The DNA sequence reveals two uninterrupted protein-coding regions of 473 (44 kd chlorophyll a apoprotein) and 353 triplets ("32 kd"-like protein). The latter gene is strikingly similar to the gene for the herbicide-binding "32 kd" protein which maps some 30 kbp distant on the plastid chromosome. The two genes overlap by 50 base pairs but are read in different phases. They may be contranscribed and the RNA modified to give several discrete species ranging in size from 1.6 to 4.6 kb. A presumptive promoter site was only identified for the "32 kd"-like protein, while potential ribosome binding and transcription termination sites are found preceding and following both genes, respectively. The polypeptides possess a high content of hydrophobic amino acids, most of which appear to be clustered in transmembrane spans. The molecular weights of 51,785 (44 kd chlorophyll a apoprotein) and 39,465 ("32 kd"-like protein) derived from the deduced amino acid sequences are higher than the experimentally determined protein sizes. Amino acid codon usage for both genes is highly selective. Comparison of the chlorophyll a apoproteins of spinach reveals regions of sequence homology.
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Several cDNA clones encoding the complete subunit II and III precursor polypeptides of the photosystem I reaction center were isolated from two spinach lambda gt1 1 expression libraries by immunoscreening and homologous hybridization. The identity of the recombinant cDNAs was verified by an N-termin