Hanisch, A., Holder, M.V., Choorapoikayil, S., Gajewski, M., Ozbudak, E.M., and Lewis, J. (2013) The elongation rate of RNA polymerase II in zebrafish and its significance in the somite segmentation clock. Development (Cambridge, England). 140(2):444-453.
A gene expression oscillator called the segmentation clock controls somite segmentation in the vertebrate embryo. In zebrafish,
the oscillatory transcriptional repressor genes her1 and her7 are crucial for genesis of the oscillations, which are thought to arise from negative autoregulation of these genes. The
period of oscillation is predicted to depend on delays in the negative-feedback loop, including, most importantly, the transcriptional
delay – the time taken to make each molecule of her1 or her7 mRNA. her1 and her7 operate in parallel. Loss of both gene functions, or mutation of her1 combined with knockdown of Hes6, which we show to be a binding partner of Her7, disrupts segmentation drastically. However,
mutants in which only her1 or her7 is functional show only mild segmentation defects and their oscillations have almost identical periods. This is unexpected
because the her1 and her7 genes differ greatly in length. We use transgenic zebrafish to measure the RNA polymerase II elongation rate, for the first
time, in the intact embryo. This rate is unexpectedly rapid, at 4.8 kb/minute at 28.5°C, implying that, for both genes, the
time taken for transcript elongation is insignificant compared with other sources of delay, explaining why the mutants have
similar clock periods. Our computational model shows how loss of her1 or her7 can allow oscillations to continue with unchanged period but with reduced amplitude and impaired synchrony, as manifested
in the in situ hybridisation patterns of the single mutants.