Single mutations in sasA enable a simpler ΔcikA gene network architecture with equivalent circadian properties.
The circadian input kinase of the cyanobacterium Synechococcus elongatus PCC 7942 (CikA) is important both for synchronizing circadian rhythms with external environmental cycles and for transferring temporal information between the oscillator and the global transcriptional regulator RpaA (regulator of phycobilisome-associated A). KOs of cikA result in one of the most severely altered but still rhythmic circadian phenotypes observed. We chemically mutagenized a cikA-null S. elongatus strain and screened for second-site suppressor mutations that could restore normal circadian rhythms. We identified two independent mutations in the Synechococcus adaptive sensor A (sasA) gene that produce nearly WT rhythms of gene expression, likely because they compensate for the loss of CikA on the temporal phosphorylation of RpaA. Additionally, these mutations restore the ability to reset the clock after a short dark pulse through an output-independent pathway, suggesting that SasA can influence entrainment through direct interactions with KaiC, a property previously unattributed to it. These experiments question the evolutionary advantage of integrating CikA into the cyanobacterial clock, challenge the conventional construct of separable input and output pathways, and show how easily the cell can adapt to restore phenotype in a severely compromised genetic network.
Center for Circadian Biology and Kavli Institute for Brain and Mind, University of California, San Diego, La Jolla, CA 92093-0116.
Ryan K
RK
Shultzaberger
Center for Circadian Biology and.
Joseph S
JS
Boyd
Kavli Institute for Brain and Mind, University of California, San Diego, La Jolla, CA 92093-0116.
Takeo
T
Katsuki
Center for Circadian Biology and sgolden@ucsd.edu rgreenspan@ucsd.edu.
Susan S
SS
Golden
Center for Circadian Biology and Kavli Institute for Brain and Mind, University of California, San Diego, La Jolla, CA 92093-0116 sgolden@ucsd.edu rgreenspan@ucsd.edu.
Ralph J
RJ
Greenspan
D001426:Bacterial Proteins / Q000235:genetics*; D002940:Circadian Rhythm; D053263:Gene Regulatory Networks; D005798:Genes, Bacterial; D017354:Point Mutation; D011110:Polymorphism, Genetic; D011494:Protein Kinases / Q000235:genetics*; D046940:Synechococcus / Q000235:genetics*
D001426:Bacterial Proteins; D011494:Protein Kinases; C411989:CikA protein, bacteria
cyanobacteria; evolution; gene network