A genetic locus targeted to the nuclear periphery in living cells maintains its transcriptional competence.
The peripheral nuclear lamina, which is largely but not entirely associated with inactive chromatin, is considered to be an important determinant of nuclear structure and gene expression. We present here an inducible system to target a genetic locus to the nuclear lamina in living mammalian cells. Using three-dimensional time-lapse microscopy, we determined that targeting of the locus requires passage through mitosis. Once targeted, the locus remains anchored to the nuclear periphery in interphase as well as in daughter cells after passage through a subsequent mitosis. Upon transcriptional induction, components of the gene expression machinery are recruited to the targeted locus, and we visualized nascent transcripts at the nuclear periphery. The kinetics of transcriptional induction at the nuclear lamina is similar to that observed at an internal nuclear region. This new cell system provides a powerful approach to study the dynamics of gene function at the nuclear periphery in living cells.
Kumaran
R Ileng
RI
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Spector
David L
DL
D002460:Cell Line; D002875:Chromosomes / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007399:Interphase; D007700:Kinetics; D034882:Lamins / Q000032:analysis / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis; D008938:Mitosis / Q000502:physiology; D008957:Models, Genetic; D034881:Nuclear Lamina / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D014158:Transcription, Genetic / Q000502:physiology*
D034882:Lamins; D008164:Luminescent Proteins