Preparation and crystallization of RNA
1Department of Biochemistry, Purdue University, West Lafayette, IN, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 3, 2007
Summary
Advances in RNA synthesis, cryocrystallography, and synchrotron X-ray sources enable detailed structural studies of RNA molecules, from simple to complex forms.
Area of Science:
- Structural Biology
- Biochemistry
- Molecular Biology
Background:
- RNA structure determination has advanced significantly due to new technologies.
- Crystallography is a key method for elucidating RNA structures.
- Previous limitations in RNA crystallography have been largely overcome.
Purpose of the Study:
- To present protocols for RNA synthesis, purification, crystallization, and derivatization.
- To facilitate the study of diverse RNA structures using X-ray crystallography.
- To contribute to the growing database of RNA structural information.
Main Methods:
- Large-scale in vitro RNA synthesis (chemical and enzymatic).
- Cryocrystallography techniques for handling sensitive RNA crystals.
- Utilization of high-intensity synchrotron X-ray beamlines.
Main Results:
- Milligram quantities of RNA are now routinely synthesized.
- RNA crystals, though radiation-sensitive, can be studied effectively.
- A comprehensive database of RNA structures, including complex molecules like ribosomes, is now available.
Conclusions:
- Modern crystallographic methods have revolutionized RNA structure analysis.
- Protocols for RNA preparation and crystallization are essential for structural studies.
- The ability to solve complex RNA structures has expanded our understanding of molecular biology.
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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,...
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01:10Transcription
Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
01:17Transcription
Transcription is the synthesis of RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in correctly synthesizing messenger RNA (mRNA). Transcriptional regulation is responsible for the differentiation of different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds of RNA Molecules
In eukaryotes,...
Transcription Can Produce Different Kinds of RNA Molecules
In eukaryotes,...
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...
01:20Types of RNA
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RNA Performs Diverse...
RNA Performs Diverse...