Iron-dependent protection of the Synechococcus ferredoxin I transcript against nucleolytic degradation requires cis-regulatory sequences in the 5' part of the messenger RNA.
We have previously reported that the ferredoxin I gene from Synechococcus sp. PCC 7942 is regulated by iron at the level of differential mRNA stability. To identify iron-responsive elements in the Synechococcus ferredoxin transcript, we have tested chimaeric constructs containing translational fusions between the Synechococcus and the Anabaena sp. PCC 7937 ferredoxin genes for iron-dependent expression in transgenic Synechococcus strains. This strategy was based on the observation that the level of the Anabaena ferredoxin mRNA did not increase upon iron addition in Synechococcus. Our results show that the presence of the first 207 nucleotides of the Synechococcus ferredoxin transcript is sufficient to confer iron responsiveness to the chimaeric transcripts. This iron responsiveness was accomplished by an increased stability of the chimaeric transcript in the presence of iron, as was found for the intact Synechococcus ferredoxin gene. Addition of the translation inhibitor chloramphenicol to the cultures led to a rapid stabilization, in low- and high-iron conditions, of the wild-type Synechococcus ferredoxin transcript as well as all chimaeric ferredoxin transcripts tested. These results suggest the existence of a constitutively expressed nuclease capable of degrading the ferredoxin transcripts. They further support the suggestion that the first 207 nucleotides of the Synechococcus transcript contain a specific sequence that is recognized by an iron-responsive factor and that this interaction leads to protection against degradation.
Bovy
A
A
Department of Molecular Cell Biology, University of Utrecht, Netherlands.
de Kruif
J
J
de Vrieze
G
G
Borrias
M
M
Weisbeek
P
P
D017033:Anabaena / Q000235:genetics; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002701:Chloramphenicol / Q000494:pharmacology; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics*; D017931:DNA Primers; D005288:Ferredoxins / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D012260:Ribonucleases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014158:Transcription, Genetic
D017931:DNA Primers; D005288:Ferredoxins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D002701:Chloramphenicol; D007501:Iron; D012260:Ribonucleases; D005966:Glucuronidase