A light-repressed transcript found in Synechococcus PCC 7002 is similar to a chloroplast-specific small subunit ribosomal protein and to a transcription modulator protein associated with sigma 54.
The gene encoding a novel light-repressed transcript (lrtA) contained within a 2.7-kbp EcoRI fragment has been cloned and sequenced from the unicellular cyanobacterium, Synechococcus PCC 7002. Northern analysis indicates that this transcript is synthesized rapidly in the dark, but upon 20 min of illumination, transcript levels fall below detectable limits. An open reading frame was located 378 bases from the start of the transcript which encodes a 21-kDa protein with significant homology to two hitherto different proteins. The protein sequence LrtA showed 37% sequence identity and 58% sequence similarity to the chloroplast-specific small subunit ribosomal protein, S30, and 37% sequence identity and 60% sequence identity and 60% sequence similarity to the reported transcription modulator protein of sigma 54 found in Klebsiella pneumonia and Azotobacter vinelandii. Expression of the lrtA gene product is not detectable within 1 h after placing the cells in the dark, however, within 2.5 min of illumination, [35S]methionine incorporated into a 21-kDa protein. To a lessor extent, [35S]methionine incorporation into a 17- and a 14-kDa protein was also seen which was followed by two other recognizable waves of translation at 5 and 10 min. This incorporation was not blocked by rifampicin added to dark-adapted cells prior to illumination. [35S]Methionine pulsed-labeling experiments suggested that the translation of lrtA occurred only during the first 10 min of reillumination of dark-adapted cells. The loss of initial [35S]methionine labeling in the light of the 21-kDa protein in a kanamycin-interrupted lrtA gene mutant suggests that the lrtA codes for the 21-kDa protein.
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Department of Biochemical and Biophysical Sciences, University of Houston, Texas 77204-5934.
Varughese
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Widger
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism; D000458:Cyanobacteria / Q000235:genetics* / Q000528:radiation effects; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D008027:Light; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D051877:RNA Polymerase Sigma 54; D012333:RNA, Messenger / Q000528:radiation effects; D012269:Ribosomal Proteins / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D012808:Sigma Factor / Q000235:genetics*; D014158:Transcription, Genetic / Q000528:radiation effects
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D012808:Sigma Factor; C088911:lrtA protein, Synechococcus sp.; C040469:ribosomal protein S30; D012321:DNA-Directed RNA Polymerases; D051877:RNA Polymerase Sigma 54