Transcriptional Bursting in Confined Cell-Free Reactions

Patrick M. Caveney1,2, Sarah E. Norred1,2, Charles Chin1,2, Scott T. Retterer1,2, Charles P. Collier2,

Michael L. Simpson1,2,3

1Bredesen Center, University of Tennessee Knoxville, 2Center for Nanophase Materials Sciences, Oak Ridge

National Laboratory, 3Department of Materials Science and Engineering, University of Tennessee Knoxville

Abstract:

Transcriptional bursting, bursts of transcription followed by periods of inactivity, is widely observed across all life1,2,3,4,5,6,7. This phenomenon is important for many cell fate decisions such as proviral latency in HIV8,9,10,11,12. The widely studied two-state (or random telegraph) model is

13,14,15,16,17,18

characterized by the transitions between ON and OFF, kON and kOFF respectively

. Many

mechanisms have been proposed including: supercoiling19, transcription factor kinetics20, chromatin

remodeling21,22, and transcriptional reinitiation23,24,25. However, these studies neglect spatial

correlations caused by in vivo crowding (40%26) and confinement (~10 femtoliters)27,28 that may

contribute to transcriptional bursting29,30. We present experimental and simulation evidence of

transcriptional bursting in confined cell-free protein synthesis reactions absent the proposed molecular mechanisms of bursting. And demonstrate that the magnitude and dynamics of bursting may be manipulated by control of confinement.

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