THE PSBB GENE-CLUSTER OF THE CHLAMYDOMONAS-REINHARDTII CHLOROPLAST - SEQUENCE AND TRANSCRIPTIONAL ANALYSES OF PSBN AND PSBH

We have sequenced and characterized the complete psbB gene cluster of Chlamydomonas reinhardtii chloroplast DNA. Although the petB and petD genes are located elsewhere, the sequential order of psbB, ORF31, psbN and psbH is identical to that of the psbB operon in higher plants. Al so, intergenic non-coding regions are much larger in the Chlamydomonas gene cluster. Northern blot analyses indicate the formation of dicist ronic transcripts of psbB and ORF31 and monocistronic transcripts of p sbN and psbH. It is unclear whether a psbB operon is transcribed to yi eld a large polycistronic precursor but northern blot analysis with to tal RNA from cells grown at 15-degrees-C does not detect an increased complexity of the transcripts, as has been found in studies of the psb B operon of higher plants. From primer extension and nuclease protecti on assays, it is apparent that 5' and 3' processing of the primary psb H transcript results in the accumulation of a heterogenous population of mRNAs. Northern blot analyses reveal transcription of Chlamydomonas psbN and show that its mRNA is much larger than that identified in li verwort and pea. The sequence identities of the PSII-H and PSII-N poly peptides as compared to their vascular plant counterparts is 50 to 62% . While the amino acid sequences of PSII-H and PSII-N proteins are sig nificantly conserved, the mass of PSII-H from Chlamydomonas is signifi cantly larger.