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RNA Splicing

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...

pre-mRNA Processing

pre-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...

Pre-mRNA Processing

Pre-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...

RNA Splicing

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...

Nuclear Export of mRNA

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...

Nuclear Export of mRNA

Nuclear Export of mRNA

Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...

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Related Experiment Video

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Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

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A nuclear cap binding protein complex involved in pre-mRNA splicing

E Izaurralde¹, J Lewis, C McGuigan

¹European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.

|August 26, 1994

Summary

The cap-binding protein complex (CBC), composed of CBP80 and CBP20, plays a crucial role in nuclear pre-mRNA splicing. This complex is essential for recognizing pre-mRNA and facilitating early steps in splicing complex formation.

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Area of Science:

Molecular Biology
Cell Biology
Biochemistry

Background:

The cap-binding protein complex (CBC) is found in the nuclei of HeLa cells.
CBC comprises two known proteins: CBP80 and CBP20.

Purpose of the Study:

To characterize the cap-binding protein complex (CBC) and elucidate its function in pre-mRNA processing.
To investigate the role of CBC in the nuclear pre-mRNA splicing pathway.

Main Methods:

Purification of CBC from HeLa cell nuclei.
Cofractionation and coimmunoprecipitation assays to confirm CBC composition.
In vitro and in vivo splicing assays using modified cap analogs.
Analysis of pre-mRNA splicing in CBC-depleted extracts.

Main Results:

Purified CBC consists of CBP80 and CBP20, which cofractionate and are coimmunoprecipitable.
Modified cap analogs inhibit splicing and bind to CBC, suggesting a role in splicing.
CBC depletion from extracts blocks an early step in splicing complex formation with adenoviral pre-mRNA.

Conclusions:

The cap-binding protein complex (CBC) is a functional entity involved in pre-mRNA splicing.
CBC likely plays a critical role in the recognition of pre-mRNA.
CBC is essential for efficient splicing complex assembly during pre-mRNA processing.