Study shows circadianclock helps cells recover during starvation elife elife
Cultures were incubated in 2% liquid medium before the inoculation to race tubes. .
We completed this section in the Materials and methods with this information:"As levels of previously used housekeeping gene actin decreased under starvation conditions, additional genes were selected based on literature data . We found gna-3 to be the least variable under the applied experimental conditions. Cvalues are presented in Figure 1 – Source data 14. For sequences of oligonucleotides and hydrolysis probes see Key resources table.
P36 Figure 5D: The race tube method is unclear. The agar medium is not specified. How many days of growth are we looking at on the race tubes? The Y axes of the graphs should use the same scale, starting at zero and ending at the same value. To better justify the choice of different protocols , we integrated more explanation into the description of the experimental procedures in the Results section . Based on the suggestion of the reviewer we also changed the order of Figure panels. In the revised version Figure 1 shows expression of clock proteins and RNAs in both LL and DD, while Figure 2 summarizes the data concerning the light induction of frq, wc-1 and al-2 transcription and the subcellular localization of the clock components.
Page 9"As shown in Figure 3A, absence of a functional FRQ protein attenuated the effects of starvation, i.e. amount of WC-1 did not change in response to glucose deprivation, whereas WC-2 levels were moderately reduced." Isn't likely that a reduction in WC1 and WC2 levels could not be measured in response to starvation because they are both already so low in frq9 mutant? Again I feel that the evidence that FRQ participates in the starvation of WC abundance is not compelling.
I think the Discussion would be improved by a comparison of the effects of glucose on the Neurospora clock with the effects of glucose found in mice and Arabidopsis.
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