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Using the E1A Minigene Tool to Study mRNA Splicing Changes
Published on: April 22, 2021
Differential dynamics of splicing factor SC35 during the cell cycle
Kaushlendra Tripathi1, Veena K Parnaik
1Centre for Cellular and Molecular Biology, Hyderabad, India.
Journal of Biosciences
|November 14, 2008
Summary
Splicing factor SC35 dynamics change during the cell cycle. Its mobility varies across cellular compartments, notably higher in metaphase cytoplasm than in interphase speckles or mitotic granules.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Nuclear speckles are dynamic structures housing pre-mRNA splicing factors.
- These speckles disassemble during mitosis and reassemble in telophase as mitotic interchromatin granules (MIGs).
Purpose of the Study:
- To investigate the dynamic behavior and mobility of the splicing factor SC35 throughout the cell cycle.
- To understand how cellular compartmentalization influences SC35 dynamics during mitosis.
Main Methods:
- Utilized HeLa cells expressing green fluorescent protein (GFP)-SC35 for live-cell imaging.
- Employed Fluorescence Recovery After Photobleaching (FRAP) to measure protein mobility.
- Applied cyclin-dependent kinase (CDK) inhibitors to assess their impact on nuclear organization and protein dynamics.
Main Results:
- GFP-SC35 localized to speckles in interphase, dispersed in metaphase, and concentrated in telophase nuclei and MIGs.
- FRAP revealed distinct GFP-SC35 mobility in different mitotic compartments, with 3-fold higher mobility in metaphase cytoplasm compared to interphase speckles, nucleoplasm, or MIGs.
- CDK inhibition altered nuclear speckle and nucleoli organization, affecting GFP-SC35 and GFP-fibrillarin mobility.
Conclusions:
- The dynamic behavior of SC35 is significantly influenced by its localization within specific cellular compartments during the cell cycle.
- Mitotic progression and cell cycle regulation impact the mobility and organization of splicing factors.
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