STOCHASTIC GENE EXPRESSION IN A LENTIVIRAL POSITIVE FEEDBACK LOOP: HIV-1 TAT FLUCTUATIONS DRIVE PHENOTYPIC DIVERSITY Leor S. Weinberger1§*, John C. Burnett3, Jared E. Toettcher2, Adam P. Arkin2,4*‡, and David V. Schaffer3‡ Biophysics Graduate Group1, Depts. of Bioengineering2, The Howard Hughes Medical Institute2 Chemical Engineering3, and the Helen Wills Neuroscience Institute3, University of California, Berkeley, CA 94720 and Physical Biosciences Division4, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 * corresponding authors:
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[email protected], Tel: (510) 495-2116, Fax (510) 486-6129 ‡ these authors contributed equally to this work Running Title: Stochastics in HIV-1 transactivation Manuscript Information 50 text pages (abstract, text body, and references) 6 Figures, 1 Table, 13 Equations Character Count (text, figure legends, methods, references): 54,548 Abstract: 143 Words Abbreviations: LTR. Long Terminal Repeat; IRES, Internal Ribosomal Entry Site; LGIT, LTR-GFP-IRES-Tat (an HIV-1 derived lentiviral vector); LG, LTR-GFP (an HIV-1 derived lentiviral vector); ODE, Ordinary Differential Equation; HERV, Human Endogenous Retrovirus; RNAPII, RNA Polymerase II; MOI, Multiplicity of Infection; GFP, Green Fluorescent Protein; TNF!, Tumor Necrosis Factor !; PMA, Phorbol Myristate Acetate; TSA, Trichostatin A; SINE, Short Interspersed Nuclear Element; LINE, Long Interspersed Nuclear Element; PheB, Phenotypic Bifurcation; PTEFb, Positive Transcriptional Elongation Factor b; Cdk9, Cyclin dependent kinase 9; RFU, Relative Fluorescence Units. SUPPLEMENTARY INFORMATION ATTACHED. § Current address: Dept. of Molecular Biology, Princeton University, Princeton, NJ 08544- 1014 Page 1 of 35 SUMMARY HIV-1 Tat transactivation is vital for completion of the viral lifecycle and has been implicated in determining proviral latency.