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Pozhitkov, A. E.

Publications and source records attributed to Pozhitkov, A. E..

2 recordsLinked to original sources

Distinct sequence patterns in the active postmortem transcriptome

Our previous study found more than 500 transcripts significantly increased in abundance in the zebrafish and mouse several hours to days postmortem relative to live controls. The current literature suggests that most mRNAs are post-transcriptionally regulated in stressful conditions, we rationalized that the postmortem transcripts must contain sequence features (3 to 9 mers) that are unique from those in the rest of the transcriptome - specifically, binding sites for proteins and/or non-coding RNAs involved in regulation. Our new study identified 5117 and 2245 over-represented sequence features in the mouse and zebrafish, respectively. Some of these features were disproportionately distributed along the transcripts with high densities in the 3-UTR region of the zebrafish (0.3 mers/nt) and the ORFs of the mouse (0.6 mers/nt). Yet, the highest density (2.3 mers/nt) occurred in the ORFs of 11 mouse transcripts that lacked UTRs. Our results suggest that these transcripts might serve as molecular sponges that sequester RNA binding proteins and/or microRNAs, increasing the stability and gene expression of other transcripts. In addition, some features were identified as binding sites for Rbfox and Hud proteins that are also involved in increasing transcript stability and gene expression. Hence, our results are consistent with the hypothesis that transcripts involved in responding to extreme stress have sequence features that make them different from the rest of the transcriptome, which presumably has implications for post-transcriptional regulation in disease, starvation, and cancer.\n\nABBREVIATIONS

systems biology

Developing a low-cost milliliter-scale chemostat array for precise control of cellular growth

Multiplexed milliliter-scale chemostats are useful for measuring cell physiology under various degrees of nutrient limitation and for experimental evolution. In each chemostat, fresh medium containing a growth rate-limiting metabolite is pumped into the culturing chamber at a constant rate, while culture effluent exits at an equal rate. Although such devices have been developed by various labs, key parameters - the accuracy and precision of flow rate and the operational range - are not explicitly characterized. Here we report the development of multiplexed milliliter-scale chemostats where flow rates for eight chambers can be independently controlled to vary within a wide range, corresponding to population doubling times of 3~ 13 hours. Importantly, flow rates are precise and accurate without the use of expensive feedback systems. Among the eight chambers, the maximal coefficient of variation in flow rate is less than 3%, and average flow rates are only slightly below targets, i.e., 3-6% for 13-hour and 0.6-1.0% for 3-hour doubling times. This deficit is largely due to evaporation and should be correctable. We experimentally demonstrate that our device allows accurate and precise quantification of population phenotypes.

microbiology