Stochastic mRNA Synthesis in Mammalian Cells Summary

Brian Kerrigan

Biol 1615 | Wednesday 5:30 pm

Mr. Jack Later

July 19, 2017

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Introduction

Scientists have observed that genetically identical populations of bacteria and yeast can

produce differing amounts of proteins. Many have thought it is due to extrinsic factors

and others due to random internal molecular events. Recent information points to the

variability being due to intrinsic random factors. This study attempts to measure gene

expression variability from cell to cell of mammalian cells by counting single molecules

of mRNA. Precisely measuring the products of gene expression, this study shows that

genes produce on and off, intermittently. They show that the sporadic expression of

genes are intrinsically random and not due to extrinsic factors.

Materials and Methods

The scientists constructed a DNA fragment to produce a multimer protein to be

measured in the cells. Reporter genes were created to help identify the protein via

fluorescents. The reporter genes were put in expression vectors to allow for their

expression in mammalian cells. Cell lines were created from cloned cells to provide an

opportunity to have several types to compare. These cells were cultured to grow more.

Probes were constructed and used to identify the fluorescent proteins. Images were

then taken to count the mRNA transcription levels in a frame. Statistics were used to

estimate the entire cells mRNA transcription levels. mRNA decay rate was then

measured. Randomization simulators were used to process the data.

Results

Images in the report show that transcription happens in bursts because there is

significant uneven distribution of the fluorescent light, implicating that it has not diffused.
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Two cells from the same parent cell show different transcriptional behavior. This proves

that extrinsic factors such as position in the cell cycle are not the largest influence on

transcription. Increasing transcriptions factors binding sites or transcription factors

themselves, increases the the average transcription burst size. This does not affect its

frequency. Random, infrequent gene activation and inactivation events were found to

control the variability in mRNA levels, not extrinsic factors.

Discussion

mRNA levels in mammalian cells vary in both reporter genes and native genes because

of intrinsic random, seldom events of gene activation. Transcription events are

regulated by levels of activators proteins and the number of transcription factors; this

can affect areas of the genome and not just specific genes. Variations are intrinsically

random, not due to extrinsic factors. Transcriptional bursts most likely correspond to

random events of chromatin (gene) remodeling. When the gene is grouping together it

would decrease transcription, when it is unravelling it would increase transcription.

Implications of this study on cellular function suggest that proteins with a longer half-life

see smaller fluctuations and protein transcription serves to only top-off the

comparatively small loss compared to the larger presence of the protein. It is likely that

essential genes which are transcribed in in bursts should have longer half-lives.

Single-cell organisms may use this oscillation in protein concentrations to survive an

unpredictable and changing environment. However, because multicellular organisms

have specialized cells the variability may be designed rather to produce a variety of

function or behavior in a tissue or organ.

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Bibliography

http://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.0040309,
visited 7/18/2017