Related Experiment Video
Updated: Jul 11, 2026
09:45
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
Sumoylation modulates the assembly and activity of the pre-mRNA 3' processing complex
Vasupradha Vethantham1, Nishta Rao, James L Manley
1Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Molecular and Cellular Biology
|October 10, 2007
Summary
Small ubiquitin-like modifier (SUMO) regulates mRNA 3'-end processing by modifying key factors like symplekin and CPSF-73. SUMOylation is crucial for the assembly and activity of the processing complex, impacting cell viability.
Area of Science:
- Molecular Biology
- Gene Expression Regulation
Background:
- Eukaryotic pre-mRNA 3'-end formation involves a complex, regulated set of factors.
- The precise mechanisms controlling this intricate process are still being elucidated.
Purpose of the Study:
- To investigate the novel role of small ubiquitin-like modifier (SUMO) in mammalian pre-mRNA 3'-end processing.
- To identify specific factors involved in 3'-end formation that are regulated by SUMOylation.
Main Methods:
- Identification of symplekin and CPSF-73 as SUMO modification substrates.
- Determination of conserved sumoylation sites in symplekin and CPSF-73.
- Functional analysis using sumoylation-deficient mutants and manipulation of sumoylation status in nuclear extracts (SUMO protease, ubc9 siRNA).
Main Results:
- Symplekin and CPSF-73 were confirmed as SUMO targets with conserved sumoylation sites.
- A sumoylation-deficient symplekin mutant impaired cell viability rescue.
- Altering sumoylation status inhibited 3'-end processing and impaired polyadenylation complex assembly.
Conclusions:
- Small ubiquitin-like modifier (SUMO) plays a critical role in regulating mammalian mRNA 3'-end processing.
- SUMOylation of symplekin and CPSF-73 enhances the assembly and activity of the 3'-end-processing complex.
More Related Videos
Related Concept Videos
02:41Chromatin Structure Regulates pre-mRNA Processing
In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
The chromatin structure, especially...
02:01pre-mRNA Processing
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
02:01Pre-mRNA Processing
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
01:35Pre-mRNA Processing: Modification of pre-mRNA Ends
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...
01:23Regulation of Expression at Multiple Steps
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
02:53Ribosomal RNA Synthesis
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...