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Related Experiment Videos

Nuclear organization and gene expression

D L Spector1

  • 1Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York, 11724, USA. spector@cshl.org

Experimental Cell Research
|December 15, 1996
PubMed
Summary
This summary is machine-generated.

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Splicing factors in cells are found in a speckled pattern, moving to storage sites when transcription or splicing is inhibited and returning to active sites when transcription resumes. This localization reflects cellular transcriptional and splicing activity.

Area of Science:

  • Cellular biology
  • Molecular genetics
  • Biochemistry

Background:

  • Pre-mRNA splicing factors exhibit a speckled localization in actively transcribing cells.
  • This pattern corresponds to interchromatin granule clusters and perichromatin fibrils.
  • Transcription is believed to occur at perichromatin fibrils, not interchromatin granule clusters.

Purpose of the Study:

  • To investigate the dynamic localization of splicing factors in relation to cellular transcription and pre-mRNA splicing activity.
  • To determine the functional significance of interchromatin granule clusters and perichromatin fibrils in splicing factor organization.

Main Methods:

  • Inhibition of RNA polymerase II transcription and pre-mRNA splicing.
  • Introduction of exogenous DNA templates into cell nuclei.

Related Experiment Videos

  • Fluorescence and electron microscopy to observe factor localization.
  • [3H]uridine incorporation studies to assess transcription sites.

    • Main Results:

    • Splicing factors redistribute to interchromatin granule clusters upon inhibition of transcription or splicing.
    • Interchromatin granule clusters enlarge and become more uniform in shape under these conditions.
    • Exogenous DNA templates recruit splicing factors to new transcription sites.

    Conclusions:

    • Splicing factors are stored/reassembled in interchromatin granule clusters and recruited to active transcription sites (perichromatin fibrils).
    • The speckled localization of splicing factors is a direct indicator of active transcription and pre-mRNA splicing within the cell.