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Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
Published on: April 26, 2017
The splicing factor SF2/ASF binds to ARS homologs in a human rDNA replication origin
Mai He1, Dimal Shah, Hae Yoon Grace Choung
1Department of Pathology and Laboratory Medicine, UMDNJ-New Jersey Medical School, Newark, NJ 07103, USA.
Cell Cycle (Georgetown, Tex.)
|July 28, 2009
Summary
Human ribosomal DNA replication origins bind alternate splicing factor SF2/ASF and GAPDH. This SF2/ASF interaction in cell nuclei suggests a new role in oncogenesis and cancer cell transformation.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- DNA replication origins in yeast (Saccharomyces cerevisiae) require origin replication complex (ORC) proteins and ARS consensus sequences (ACS).
- Mammalian replication origins lack conserved ACS elements, showing high sequence variability despite conserved ORC proteins.
- Previously identified human ribosomal DNA replication initiation sites contain ACS homologues within DNA unwinding elements and matrix attachment regions.
Purpose of the Study:
- To investigate protein binding to ACS homologues within human ribosomal DNA replication initiation sites.
- To determine if SF2/ASF and GAPDH bind these sequences in vitro.
- To provide evidence for the in vivo interaction of SF2/ASF within intact cell nuclei.
Main Methods:
- In vitro protein binding assays to assess interactions with ACS homologues.
- Cellular localization studies to confirm in vivo SF2/ASF binding within nuclei.
Main Results:
- Human ribosomal DNA ACS homologues specifically bind the alternate splicing factor SF2/ASF and GAPDH in vitro.
- Evidence was presented demonstrating that SF2/ASF interacts with these sites within the nuclei of intact cells.
Conclusions:
- The binding of SF2/ASF to human ribosomal DNA replication origins suggests a novel role for this splicing factor.
- This interaction may represent an additional mechanism by which SF2/ASF contributes to oncogenesis and the transformed cell phenotype, given its link to alternatively spliced regulatory proteins.
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