title: Combining protein and mRNA quantification to decipher transcriptional regulation.
abstract: We combine immunofluorescence and single-molecule fluorescence in situ hybridization (smFISH), followed by automated image analysis, to quantify the concentration of nuclear transcription factors, number of transcription factors bound, and number of nascent mRNAs synthesized at individual gene loci. A theoretical model is used to decipher how transcription factor binding modulates the stochastic kinetics of mRNA production. We demonstrate this approach by examining the regulation of hunchback in the early Drosophila embryo.
authors.affiliation: 1] Verna &Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA. [2] Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [3] Center for Theoretical Biological Physics, Rice University, Houston, Texas, USA.
authors.forename: Heng
authors.initials: H
authors.lastname: Xu
authors.affiliation: 1] Verna &Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA. [2] Center for Theoretical Biological Physics, Rice University, Houston, Texas, USA.
authors.forename: Leonardo A
authors.initials: LA
authors.lastname: Sepúlveda
authors.affiliation: Integrative Molecular and Biomedical Sciences Graduate Program, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Figard
authors.affiliation: Verna &Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Anna Marie
authors.initials: AM
authors.lastname: Sokac
authors.affiliation: 1] Verna &Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA. [2] Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. [3] Center for Theoretical Biological Physics, Rice University, Houston, Texas, USA.
authors.forename: Ido
authors.identifier: 0000000243084959
authors.initials: I
authors.lastname: Golding
mesh_terms: D053959:Alpha-Amanitin / Q000737:chemistry; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003433:Crosses, Genetic; D004247:DNA / Q000737:chemistry; D029721:Drosophila Proteins / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D007091:Image Processing, Computer-Assisted; D017404:In Situ Hybridization, Fluorescence; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008962:Models, Theoretical; D016011:Normal Distribution; D010363:Pattern Recognition, Automated; D011506:Proteins / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D013045:Species Specificity; D013269:Stochastic Processes; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D053959:Alpha-Amanitin; D029721:Drosophila Proteins; D011506:Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D004247:DNA