Name: pre-mrna splicing and intron removal
Uploaded: 2023-04-30T18:28:44.000Z
Duration: 1 min 32 s
Channel: JoVE

K12 Standards

Virginia  Standards of Learning

HIgh School

Biology

BIO.2

BIO.2.d

Pre-mRNA Processing: RNA Splicing

When pre-mRNA is transcribed from DNA, it contains exons, protein-coding sequences, and introns, non-coding regions. RNA splicing removes the introns and attaches the exons together.
This is catalyzed at the spliceosome, a large assembly of small nuclear ribonucleoproteins, or snRNPs &#8212; complexes made up of small nuclear RNAs and proteins.
The spliceosome is composed of 5 snRNPs, U1, U2, U4, U5 and U6, and several other proteins.
First, U1 snRNP binds the 5&#39; splice site while the branch point sequence containing A is recognized by other proteins and later replaced by U2 snRNP.&#160;
Next, a complex of U4/U6 and U5 snRNPs binds to the 5&#39; splice junction and assists in bringing the 5&#8217; end to the branch point A. A transesterification reaction between the branch point A and the 5&#39; splice site creates a loop called a lariat.
Following this, another transesterification reaction occurs between the 3&#39; end of the upstream exon and the 5&#39; end of the next exon. This cleaves off the lariat containing the intron, leaving the exons attached to each other.&#160;

pre-mrna processing: 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 together.
Splicing is Mediated by the Spliceosome
Splicing occurs in the nucleosome and is mediated by a complex of proteins and small RNAs called small nuclear ribonucleoproteins (snRNPs). snRNPs, together with other proteins, form the spliceosome, which recognizes specific nucleotide sequences at the ends of the exon and intron. First, it binds to a GU-containing sequence at the 5&#39; end of the intron and to a branch point sequence containing an A towards the 3&#39; end of the intron. In a number of carefully-orchestrated steps, other snRNPs then bring the branch point close to the 5&#39; splice site. Subsequently, a chemical reaction cleaves the 5&#39; end of the intron from its upstream exon and attaches it to the branch point, forming a loop called a lariat. To release the lariat, the AG-containing sequence of the intron near the 5&#39; end of the downstream exon reacts with the 3&#39; end of the upstream exon. This reaction patches the two exons together, concluding the splicing process.
Splicing Allows the Expression of Several Proteins from a Single Gene
Typically, exons are joined together in the order in which they appear in a gene. However, during alternative splicing, different combinations of exons in pre-mRNA are combined to form mature mRNA. This produces several different proteins from a single pre-mRNA transcript.
Different patterns of alternative splicing include exon skipping, alternative 5&#39; or 3&#39; splice sites, and intron retention. These patterns are guided by the length of exons or introns and the strength of the splicing signal at the splice sites. Because of this, exons that are shorter than other exons may be overlooked by the spliceosome and omitted from the mature mRNA. In contrast, introns that are significantly shorter than other introns may evade removal by the spliceosome and are retained in the mature mRNA. As a result, alternative splicing generates variants of mature mRNA that were copied from the same stretch of DNA. The RNA sequence variants produce different proteins with additional or fewer amino acids, shifts in the reading frame, or a premature stop codon. These protein isoforms have different biological properties, including function, cellular localization, and interaction with other proteins, thereby playing a vital role in tissue- and environment-specific gene expression.
Abnormal Splicing Can Cause Diseases
Errors in splicing can produce aberrant protein isoforms, which may contribute to diseases, including cancer. For instance, alternative splicing of the BCL2L1 gene generates a long and short protein isoform&#8212;BCL-XL and BCL-XS, respectively&#8212; through the use of alternative 5&#39; splice sites. The longer BCL-XL isoform promotes cell survival and is highly expressed in several types of cancers (e.g., blood, breast, and liver cancers). Expression of the short BCL-XS isoform that promotes cell death is suppressed in cancer.

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called ...

Education

JoVE Core

Cell Biology

Unit 1

Transcription: DNA to RNA

KEY TERMS AND CONCEPTS

pre-mrna splicing and intron removal

Pre-mRNA splicing removes introns from a pre-mRNA molecule and joins the remaining exons together. This step is part of pre-mRNA processing, which helps turn the first RNA copy into a mature message that can be used later in gene expression.

Splicing is carried out by the spliceosome, a large RNA-protein complex. The spliceosome recognizes the intron boundaries and cuts out the intron. It then links the exons together in the correct order.

A key feature of splicing is the lariat, a looped RNA structure formed when the intron is removed. The intron is released in this looped form after the spliceosome completes the cut-and-join process. This mechanism ensures that the final mRNA contains only the coding regions needed for translation.

Pre-mRNA Splicing and Intron Removal

what is gene expression?

How Cells Control Gene Expression

Cells control gene expression to decide when a gene makes a protein. A gene is a stretch of DNA that holds the instructions for functional RNAs and proteins. Because DNA is built from nucleotides and proteins are built from amino acids, messenger RNA (mRNA) acts as the link between them. The first step is transcription. During transcription, mRNA copies the DNA blueprint. In eukaryotes, this happens in the nucleus through complementary base-pairing with the DNA template. The mRNA is then...

rna structure

RNA Shapes and Protein Synthesis

RNA shapes and protein synthesis are linked through the molecule’s single-stranded structure. RNA is made of ribonucleotides joined by phosphodiester bonds, and it can fold into secondary and tertiary shapes that help control transcription and translation. These shapes are important in both regulation and protein production. Messenger RNA, transfer RNA, and ribosomal RNA are the three main RNA types used in protein synthesis. They all share the same basic build: a single strand of nucleotides.

types of rna

RNA Roles in Protein Synthesis

RNA plays key roles in protein synthesis, and its main forms work together to make proteins in the cell. Messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA) each have a specific job. RNA can also be grouped as protein-coding or non-coding RNA. mRNA is the protein-coding RNA. It carries codons, which are sets of three nucleotides that specify an amino acid. tRNA and rRNA are non-coding RNAs. tRNA acts as an adaptor that reads the mRNA sequence and brings amino acids into the...

transcription

RNA Synthesis and Gene Control

Transcription is the process that makes RNA from a DNA sequence. RNA polymerase carries out this synthesis. It is the first step in turning a gene into a protein, and several other proteins and control sequences help make messenger RNA, or mRNA, correctly. In eukaryotes, DNA is first copied into pre-mRNA. This RNA can then be processed into mature mRNA, which serves as the template for protein synthesis. In prokaryotes, translation into a polypeptide can start before transcription has finished.

bacterial rna polymerase

Bacterial RNA Polymerase and Sigma Factor

Bacterial RNA polymerase carries out transcription, the process of copying DNA into RNA. It is the enzyme that starts RNA synthesis in bacteria and helps control which genes are used. The core enzyme works with a sigma factor, which is a protein that helps the polymerase find the right place to begin. Together, they form the holoenzyme. The sigma factor is important for promoter recognition, so the enzyme can bind near the start of a gene. Once transcription begins, RNA polymerase separates...

eukaryotic rna polymerases

Three Eukaryotic RNA Polymerases

Eukaryotic RNA polymerases are the enzymes that make RNA from DNA in cells with a nucleus. In eukaryotes, this work is divided among three main RNA polymerases. Each enzyme has a specific role in transcription, the process of copying genetic information into RNA. RNA polymerase I makes most ribosomal RNA, or rRNA. RNA polymerase II produces messenger RNA, or mRNA, which carries the instructions for making proteins. RNA polymerase III makes transfer RNA, or tRNA, and also some small RNAs used...

general transcription factors

Tissue-Specific Transcription Factors in Gene Control

Tissue-specific transcription factors help control which genes turn on in different mammal cells. These proteins bind to promoter sequences and can direct a gene to be expressed in one tissue but not another. This selective control helps cells carry out very different jobs in the body. A clear example is the beta globin gene, which makes part of hemoglobin. The gene is present in all body cells, but it is expressed only in red blood cells. That happens because the transcription factors needed...

rna polymerase ii accessory proteins

RNA Polymerase II Helper Proteins

RNA polymerase II helper proteins support the transcription process in eukaryotic cells. They work with RNA polymerase II, the enzyme that makes RNA from a DNA template. These proteins help the enzyme start, move, and finish transcription in the right way. Some helper proteins act as accessory factors. They can assist RNA polymerase II at different stages of gene expression. Their roles are important because transcription must be controlled accurately for cells to make the correct RNA...

transcription elongation factors

RNA Polymerase Pausing and Elongation

RNA polymerase pausing and elongation factors help control how RNA is made from DNA. During transcription, RNA polymerase moves along the gene and builds an RNA strand. Elongation factors support this process by helping the enzyme continue after it pauses. These factors are part of the elongation stage of transcription. They are not the same as the proteins used to start transcription. Their role is to keep RNA polymerase moving so RNA can be copied efficiently. In high school biology, this...

pre-mrna processing: modification of pre-mrna ends

pre-mrna-processing-modification-of-pre-mrna-ends

How Pre-mRNA Becomes Mature mRNA

Pre-mRNA becomes mature mRNA through a series of processing steps in eukaryotic cells. RNA polymerase first makes the transcript, and the RNA is modified before it leaves the nucleus. The unprocessed form is called precursor mRNA, or pre-mRNA, to separate it from mature mRNA. After about 20 to 40 ribonucleotides have been added, enzymes place a cap on the 5' end of the growing RNA. This cap replaces the 5' phosphate with modified guanosine, which carries a methyl group called 7-methyl...

Pre-mRNA splicing removes introns from a pre-mRNA molecule and joins the remaining exons together. This step is part of pre-mRNA processing, which helps turn the first RNA copy into a mature message that can be used later in gene expression. Splicing is carried out by the spliceosome, a large RNA-protein complex. The spliceosome recognizes the intron boundaries and cuts out the intron. It then links the exons together in the correct order. A key feature of splicing is the lariat, a looped RNA...

chromatin structure and rna splicing

Chromatin and RNA Splicing Control

Chromatin and RNA splicing work together to shape how genes are used in a cell. Chromatin is the DNA-protein material that packages genetic information, and RNA splicing is a step in which introns are removed from a pre-mRNA transcript. The video explains how chromatin structure can influence this RNA processing step. A compact chromatin structure can make a gene less accessible, while a more open structure can make it easier for the cell to use that gene. This change in access can affect how...

alternative rna splicing

RNA Splicing in Gene Expression

RNA splicing in gene expression helps cells turn one gene into different RNA messages. During RNA processing, introns are removed and exons are joined together. This editing step changes which parts of the RNA stay in the final message. Alternative splicing gives cells a way to make more than one protein from the same gene. By including or skipping different exons, a cell can produce different RNA versions from one starting transcript. This process adds flexibility to gene expression without...

nuclear export of mrna

How mRNA Leaves the Nucleus

Messenger RNA, or mRNA, must move from the nucleus to the cytoplasm before it can be used to make protein. This step is called nuclear export of mRNA. It connects gene expression in the nucleus with protein production in the cytoplasm. During export, the cell selects mRNA molecules that are ready to leave the nucleus. These molecules pass through nuclear pores, which are openings in the nuclear envelope. The process helps keep mRNA transport organized and controlled. Once mRNA reaches the...

transfer rna synthesis

tRNA Genes and RNA Processing

Transfer RNA synthesis begins with transcription of tRNA genes into a primary RNA copy. That first copy is not yet ready to function in protein building. It must be processed before it can become mature transfer RNA. During processing, the cell trims the ends of the RNA and removes extra segments when needed. The transcript also shows that some transfer RNAs carry modified bases. These changes help create the final tRNA structure. Mature transfer RNA folds into a shape that can work in...

ribosomal rna synthesis

Ribosomal RNA and Ribosome Formation

Ribosomal RNA synthesis is part of how cells build ribosomes. Ribosomes are the structures that help make proteins. In this process, cells produce ribosomal RNA, or rRNA, which becomes a key part of the ribosome. The transcript focuses on the making of rRNA and its role in ribosome formation. Ribosomal RNA is not just a message like messenger RNA. It is a structural and functional part of the ribosome itself. Because of that, rRNA synthesis is closely tied to the assembly of ribosomes. This...

the nucleolus

Nucleolus and Ribosome Assembly

The nucleolus is a small structure inside the nucleus. It is the site where ribosome assembly begins. Ribosomes are the cell structures that help make proteins. The nucleolus forms around nucleolar organizer regions, or NORs. These regions contain the genes used to make ribosomal RNA, also called rRNA. Inside the nucleolus, rRNA is produced and combined with proteins to start building ribosomes. Because of this role, the nucleolus is closely tied to protein production in the cell. Its work...

additional subnuclear structures

Subnuclear Bodies and Their Roles

Subnuclear structures are specialized bodies inside the nucleus. They help organize nuclear activity and appear as distinct regions under the microscope. The nucleolus is one of the best-known subnuclear structures. It is the site where ribosomes begin to assemble. Cajal bodies are another nuclear structure. They are linked to the processing and assembly of RNA-related particles. These additional subnuclear structures show that the nucleus is not a uniform space. Instead, it contains...

key terms and concepts

Learn how pre-mRNA splicing removes introns, joins exons, and uses the spliceosome and lariat structure to make mature mRNA for gene expression.

Pre-mRNA Splicing and Intron Removal | JoVE K12

cell biology

JoVE Core Cell Biology illustrates complex cellular processes in easy-to-understand yet comprehensive animated video lessons. This collection focuses on the essential components and functions of cells, including organelles and membranes, cellular transport, and cell energetics. It is complemented by scientist-in-action videos that demonstrate the application of cell biology in real-world research experiments.

Minnesota Academic Standards in Science

9-12

Life Science

3.2.1 9L.3.2.1.1

Alabama Course of Study Science

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms: Structures and Processes

9-12.B.1.2

Florida Department of Education

High School

SC.912.L.16

SC.912.L.16.5

SC.912.L.16.6

south-carolina-college-and-career-ready-science-standards

South Carolina College and Career Ready Science Standards

B-LS1-1.

myJoVE Corporation

Functions of Life

Human life is characterized by a variety of functions that are essential for survival and well-being. These functions include metabolism, movement, ...

Requirements for Human Life

The Earth and its atmosphere have provided humans with air, water, and food, but these are not the only requirements for survival. Humans also require a ...

What is Homeostasis?

Maintaining homeostasis requires that the body continuously maintain its internal conditions. Each physiological condition has a particular set point, ...

Negative and Positive Feedback

Homeostatic Imbalance

Homeostasis is the maintenance of a stable internal environment within the body, which is crucial for the proper functioning of cells, tissues, organs, ...

Role of Water in Human Biology

Water is the one of the most significant components of the human body; it plays a crucial role in several physiological activities because of its unique ...

Introduction to Electrolytes

In humans, electrolytes play a vital role in various physiological processes. Balancing electrolyte levels is essential for normal body functions; their ...

pH Homeostasis

Acid-base homeostasis is essential for maintaining normal physiological activities in humans. The pH of various body fluids is strictly regulated because ...

The student will investigate and understand that chemical and biochemical processes are essential for life. Key ideas include water chemistry has an influence on life processes;

BIO.2.a

Compounds Essential to Human Function

The human body is composed of cells that are fundamentally made up of several different molecules. These molecules are essential to carry out all ...

Macromolecules

Biomolecules
Organisms contain a wide variety of organic molecules with numerous functions which depend on the chemical structures and properties of these ...

Solution Preparation
ExpandCAUTION: It is important to always use care when handling reagents. For example, iodine will stain clothes and skin so wear ...

Identifying Macromolecules with Indicator Solutions
ExpandHYPOTHESES: In this activity, the experimental hypothesis might be that the addition of an ...

pH Regulation in Cells

pH plays a critical role in maintaining normal cellular activities. It helps maintain the structure and function of various proteins, dictates the charge ...

Chemistry of the Cell

Overview of Functional Groups

Functional groups are a group of atoms with characteristic properties, which when linked to the carbon skeleton of a molecule, alter the properties of ...

Introduction to Carbohydrates

Carbohydrates, proteins, and fats are the primary macronutrients in the human diet. However, carbohydrates are the most favored source of energy in the ...

Carbohydrate Metabolism

Sugars as Energy Storage Molecules

What are Lipids?

lipid-derived-compounds-in-the-human-body

Lipid-derived Compounds in the Human Body

Fats and lipids are crucial components in the human body. Some lipid-derived compounds, such as fat-soluble vitamins, eicosanoids, lipoproteins, and ...

Fats as Energy Storage Molecules

What are Proteins?

Protein Organization

Globular Proteins

In organisms, proteins are the most abundant macromolecules. They act as the building blocks of life and play various crucial roles in the body. Proteins ...

Fibrous Proteins

Fibrous proteins are either long and narrow proteins or assemble to form long and thin structures. They contain repetitive units and usually consist of ...

Protein Denaturation

The function of proteins depends on their native three-dimensional structure, which is dictated by the amino acid sequence of the specific protein. ...

Role of Proteins in the Human Body

Proteins are the building block of life. They are also&#160; the most abundant macromolecules with as many diverse roles in the body. They are part of ...

Nucleic Acids and Nucleotides

Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell&#39;s genetic blueprint and have instructions for its ...

Nucleic Acid Structure

The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group ...

ATP Energy Storage and Release

ATP and Macromolecule Synthesis

Biological macromolecules are organic compounds, predominantly composed of carbon atoms. The carbon atoms are covalently bonded with hydrogen, oxygen, ...

The student will investigate and understand that chemical and biochemical processes are essential for life.Key ideas include  macromolecules have roles in maintaining life processes;

BIO.2.b

structural-organization-of-the-human-body-an-overview

Structural Organization of the Human Body: An Overview

It is convenient to consider the body&#39;s structures in terms of fundamental levels of organization that increase in complexity: subatomic particles, ...

Introduction to Enzymes

The use of enzymes by humans dates to 7000 BCE. Humans first used enzymes to ferment sugars and produce alcohol without knowing that this was an ...

Enzymes and Activation Energy

The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to ...

Introduction to Enzyme Kinetics

Enzyme kinetics studies the rates of biochemical reactions. Scientists monitor the reaction rates for a particular enzymatic reaction at various substrate ...

Turnover Number and Catalytic Efficiency

The turnover number of an enzyme is the maximum number of substrate molecules it can transform per unit time. Turnover numbers for most enzymes range from ...

Catalytically Perfect Enzymes

The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at ...

introduction-to-mechanisms-of-enzyme-catalysis

Introduction to Mechanisms of Enzyme Catalysis

For many years, scientists thought that enzyme-substrate binding took place in a simple &#34;lock-and-key&#34; fashion. This model stated that the enzyme ...

Ligand Binding and Linkage

Cooperative Allosteric Transitions

Ribozymes

The student will investigate and understand that chemical and biochemical processes are essential for life. Key ideas include  enzymes have a role in biochemical processes;

BIO.2.c

What are Cells?

Cells are the smallest and basic units of life, whether it is a single cell that forms the entire organism, e.g., in a bacterium, or trillions of them, ...

The Nucleus

The nucleus is a membrane-bound organelle that acts as a control center in a eukaryotic cell. It contains chromosomal DNA, which controls gene expression ...

The DNA Helix

Deoxyribonucleic acid, or DNA, is the genetic material responsible for passing traits from generation to generation in all organisms and most viruses. DNA ...

The Central Dogma

The central dogma explains the flow of genetic information from DNA nucleotides to the amino acid sequence of proteins.
RNA is the Missing Link Between ...

Mutations

Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations ...

DNA as a Genetic Template

Structure of a Gene

A gene is the fundamental unit of heredity. Every individual has two copies of each gene, one inherited from each parent. Although most people contain the ...

Chromosome Structure

What is Gene Expression?

A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised&#160; of nucleotides and proteins are ...

RNA Structure

The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form ...

Types of RNA

Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse ...

Transcription

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. ...

Bacterial RNA Polymerase

Eukaryotic RNA Polymerases

General Transcription Factors

Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of ...

RNA Polymerase II Accessory Proteins

Transcription Elongation Factors

The student will investigate and understand that chemical and biochemical processes are essential for life. Key ideas include protein synthesis is the process of forming proteins which influences inheritance and evolution; and

Pre-mRNA Splicing and Intron Removal

Description

Transcript

Tags

More videos by Standards