In vitro mutagenesis of binding site elements for the clock-controlled proteins CCTR and Chlamy 1.
The luciferin-binding protein (LBP) from the dinoflagellate, Gonyaulax polyedra, is regulated by a circadian clock at the translational level. A 22-nucleotide long interval in the lbp 3' untranslated region, which contains seven UG-repeats, was characterized as a circadian cis-acting element, to which a clock controlled factor (CCTR) binds. Recently we have found that the phylogenetically distant green alga, Chlamydomonas reinhardtii, contains a CCTR analog, called Chlamy 1. Here we show that the flanking nucleotides surrounding the UG-repeats are required for high binding activity of CCTR and Chlamy 1. The absence of three or more UG-repeats abolishes binding with both proteins.
Mittag
M
M
Botanisches Institut, Ludwig-Maximilians-Universität München, Germany.
Waltenberger
H
H
D000818:Animals; D056926:CLOCK Proteins; D002135:Calcium-Binding Proteins / Q000235:genetics*; D016825:Chlamydomonas reinhardtii / Q000235:genetics; D002940:Circadian Rhythm / Q000235:genetics*; D004141:Dinoflagellida / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D010802:Phylogeny; D014176:Protein Biosynthesis / Q000235:genetics; D015800:Protozoan Proteins; D015534:Trans-Activators / Q000235:genetics*
D002135:Calcium-Binding Proteins; D015800:Protozoan Proteins; D015534:Trans-Activators; C065397:luciferin-binding protein, dinoflagellate; D056926:CLOCK Proteins