Types and Functions of RNA

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Questions and Answers

What is the function of RNA polymerase during transcription?

  • Catalyzes phosphodiester bond formation between rNTPs (correct)
  • Synthesizes RNA in a 3′→5′ direction
  • Separates the RNA strands after synthesis
  • Releases ATP for energy during transcription

Which statement correctly describes the transcription bubble?

  • It involves approximately 8–10 base pairs of DNA
  • It is created when the DNA strands are separated around the transcription start site (correct)
  • It is the area where RNA polymerase dissociates from the DNA
  • It is where the nascent RNA strand is fully synthesized

What denotes the 'downstream' direction in transcription?

  • The sequence of rNTPs added to the growing RNA strand
  • The area where RNA synthesis begins
  • The 5′ end of the RNA strand being synthesized
  • The direction where RNA polymerase moves during transcription (correct)

What does the term 'upstream' indicate in the context of transcription?

<p>The direction opposite to which RNA polymerase moves (A)</p> Signup and view all the answers

What is the maximum rate of RNA synthesis by RNA polymerase at 37°C?

<p>1000-2000 nucleotides per minute (B)</p> Signup and view all the answers

How many base pairs of DNA are typically separated at the transcription start site?

<p>12–14 base pairs (C)</p> Signup and view all the answers

Which of the following is a characteristic of the elongation complex during RNA synthesis?

<p>It comprises RNA polymerase and the nascent RNA strand (A)</p> Signup and view all the answers

What is the simplest definition of a gene?

<p>A unit of DNA responsible for the synthesis of a single polypeptide chain or functional RNA (A)</p> Signup and view all the answers

What is the primary function of the 5' cap added to nascent RNAs?

<p>To mark the RNA as mRNA precursor and protect it from degradation (C)</p> Signup and view all the answers

Which RNA polymerase is specifically characterized by the presence of the carboxy-terminal repeat (CTD)?

<p>RNA Polymerase II (A)</p> Signup and view all the answers

During the elongation stage, what happens to the RNA structure that forms from termination signals?

<p>It destabilizes the polymerase’s hold on RNA (C)</p> Signup and view all the answers

What are the three major events that occur during the processing of eukaryotic pre-mRNA?

<p>5’ capping, 3’ cleavage and polyadenylation, and RNA splicing (B)</p> Signup and view all the answers

Which component is directly responsible for catalyzing the addition of the 5' cap during transcription?

<p>A dimeric capping enzyme (C)</p> Signup and view all the answers

What effect does 5' capping have on the negative elongation factor (NELF)?

<p>It leads to the release of NELF (C)</p> Signup and view all the answers

Which of the following statements about eukaryotic gene structure is correct?

<p>A gene includes the entire nucleic acid sequence necessary for functional gene product synthesis. (C)</p> Signup and view all the answers

What is indicated by the phosphorylation of the carboxy-terminal repeat (CTD) in RNA polymerase II?

<p>It is associated with elongation and activates capping enzymes. (A)</p> Signup and view all the answers

Which factor is NOT relevant to determining the melting temperature (Tm) of DNA?

<p>Length of the messenger RNA (A)</p> Signup and view all the answers

What type of bond links two amino acids together?

<p>Peptide bond (D)</p> Signup and view all the answers

What is the primary role of RNA Polymerase II in the cell?

<p>Transcription of messenger RNA (D)</p> Signup and view all the answers

Which of the following bonds connects guanine and cytosine in DNA?

<p>Hydrogen bond (B)</p> Signup and view all the answers

Which statement regarding the DNA replication fork is accurate?

<p>It is asymmetrical leading to different replication rates. (D)</p> Signup and view all the answers

What is the role of hnRNP proteins in mRNA processing?

<p>They contribute to RNA splicing or cleavage/polyadenylation. (C)</p> Signup and view all the answers

Which of the following statements accurately describes splicing in eukaryotic cells?

<p>Splicing can occur co-transcriptionally for longer transcripts. (C)</p> Signup and view all the answers

What is the average length of introns found in human genes?

<p>3500 bp (C)</p> Signup and view all the answers

What is a function of SR proteins in the splicing process?

<p>They recognize exonic splicing enhancers. (C)</p> Signup and view all the answers

How does the TATA box influence transcription initiation?

<p>It defines the start site for transcription. (B)</p> Signup and view all the answers

What distinguishes CpG islands from the TATA box in terms of function?

<p>CpG islands are less defined initiation sites compared to the TATA box. (D)</p> Signup and view all the answers

What percentage of single-base mutations that cause human genetic diseases affect exon definition?

<p>15 percent (D)</p> Signup and view all the answers

What occurs immediately following the cleavage and polyadenylation of short transcripts?

<p>Splicing occurs. (C)</p> Signup and view all the answers

What is the primary function of the 5’ cap on mRNA?

<p>To protect the 5’ end from degradation (A)</p> Signup and view all the answers

Which enzymes are involved in the capping of mRNA?

<p>Phosphatase, guanyl transferase, and methyl transferase (C)</p> Signup and view all the answers

How does alternative splicing affect fibronectin production in different cell types?

<p>Fibronectin mRNA varies due to inclusion or exclusion of specific exons. (B)</p> Signup and view all the answers

What is the role of hnRNP proteins in the processing of pre-mRNA?

<p>They increase accessibility and uniformity for interactions with RNA molecules. (B)</p> Signup and view all the answers

Which exons are included in the fibronectin mRNA produced by fibroblasts?

<p>EIIIA and EIIIB exons (A)</p> Signup and view all the answers

What is the importance of the reverse linkage formed during capping of mRNA?

<p>It is essential for cap recognition by ribosomes. (B)</p> Signup and view all the answers

What is the consequence of fibronectin lacking the EIIIA and EIIIB exons in hepatocytes?

<p>It circulates freely in the blood. (A)</p> Signup and view all the answers

What type of complex is formed with RNA molecules during their processing in the nucleus?

<p>Heterogeneous ribonucleoprotein particles (hnRNPs) (D)</p> Signup and view all the answers

What defines CpG islands within DNA?

<p>Regions with high GC content located within 100-1000 bp (A)</p> Signup and view all the answers

Which factor binds first to the TATA box during transcription initiation?

<p>TBP (A)</p> Signup and view all the answers

What is the role of the N-terminal domain of TFIIB?

<p>To help melt the template DNA during initiation (B)</p> Signup and view all the answers

Which RNA Polymerase is primarily responsible for the transcription of tRNA genes?

<p>RNA Polymerase III (C)</p> Signup and view all the answers

What is the function of the helicase activity of TFIIH during transcription initiation?

<p>To melt the DNA and form an open complex (B)</p> Signup and view all the answers

Which component is heavily involved in the formation of the pre-initiation complex?

<p>A combination of general transcription factors and RNA Polymerase II (C)</p> Signup and view all the answers

What happens to general transcription factors after transcription initiation?

<p>They primarily dissociate from the transcription complex (A)</p> Signup and view all the answers

What is the primary function of the cytoskeleton in a eukaryotic cell?

<p>To provide structural support and facilitate movement (A)</p> Signup and view all the answers

Flashcards

RNA types and functions

RNA exists in various types with specific roles. Examples include messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA).

RNA transcription

The process of creating RNA from a DNA template. It's a fundamental step in protein synthesis.

Gene (simplest definition)

A DNA segment that encodes a polypeptide chain or functional RNA.

Noncoding RNA

RNA molecules not directly used to make proteins.

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Transcription directionality

RNA polymerase moves along the template DNA strand in a 3' to 5' direction; RNA is synthesized 5' to 3'.

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Pyrophosphatase

An enzyme that removes pyrophosphate, a byproduct of RNA synthesis.

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Transcription bubble

The region of DNA unwound during transcription, a crucial step in the process.

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Upstream/Downstream

Terms used to describe the directionality of DNA sequences relative to the transcription start site.

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Transcription Initiation

The beginning of transcription, where RNA polymerase binds to the DNA and begins to synthesize RNA.

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Transcription Elongation Complex

The stable structure formed by RNA polymerase, template DNA, and nascent RNA.

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5' Capping of mRNA

A process that protects the 5' end of mRNA from degradation, aids in mRNA transport, and facilitates translation initiation.

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5' and 3' UTR

Non-coding regions of mRNA located at the 5' and 3' ends, influencing mRNA translation and stability.

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Alternative splicing

The process where different combinations of exons are joined together during mRNA processing, creating different mRNA isoforms from the same gene.

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Fibronectin

An adhesive extracellular protein that binds other proteins, facilitating cell-cell adhesion and cellular interactions with matrix.

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Heterogeneous ribonucleoprotein particles (hnRNPs)

Nuclear complexes containing hnRNA and proteins, involved in pre-mRNA processing and regulation.

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mRNA translation

The process by which the genetic information in mRNA is used to synthesize proteins.

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3'UTR mRNA localization

The 3' untranslated region of mRNA influencing the localization of mRNA to specific regions within the cytoplasm, affecting protein expression.

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RNA Polymerase

An enzyme that synthesizes RNA molecules from a DNA template.

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Transcription

The process of making an RNA copy of a DNA sequence.

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Elongation (transcription)

The stage of transcription where RNA polymerase adds nucleotides to the growing RNA chain.

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Termination signals (transcription)

DNA sequences that signal the end of transcription.

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Eukaryotic RNA Polymerases

Three different enzymes that transcribe different types of RNA in eukaryotic cells.

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Carboxy-Terminal Repeat (CTD)

An essential region in RNA polymerase II, crucial for processing and regulation.

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Eukaryotic Gene Structure

The complete DNA sequence required for making a functional gene product (protein or RNA).

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5' Capping

The addition of a special chemical structure (the cap) to the 5' end of a nascent RNA molecule.

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3’ cleavage and polyadenylation

The process of cutting and adding a tail (poly-A tail) to the 3' end of a pre-mRNA molecule.

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RNA splicing

The process of removing non-coding regions (introns) from pre-mRNA and joining coding regions (exons).

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hnRNP proteins

Proteins that interact with RNA sequences involved in splicing or cleavage/polyadenylation, contributing to the structure recognized by RNA-processing factors. They also help export mRNA from the nucleus.

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RNA splicing

The process of removing introns and joining exons in a pre-mRNA molecule to create a mature mRNA molecule.

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Spliceosome

A large ribonucleoprotein complex composed of snRNPs and other proteins that carries out RNA splicing.

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Exon

The part of a gene that codes for a protein or part of a protein.

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Intron

Non-coding regions of a gene that are removed during RNA splicing.

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Splicing occurs co-transcriptionally

Splicing happens at the same time as transcription for long transcripts. For shorter transcripts, it follows cleavage and polyadenylation.

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Splicing snRNPs

Small nuclear ribonucleoproteins that are part of the spliceosome and are essential for splicing.

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TATA box

A DNA sequence that serves as a critical promoter element for transcription start sites in eukaryotes, usually about 25-30 base pairs upstream from the transcription start site.

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Initiator Sequences

Sequences that act as alternative or supplementary promotion elements for transcription, often found near the transcription start site.

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CpG Islands

DNA regions rich in CpG dinucleotides, often found in eukaryotic promoters and associated with genes that are not required in large amounts.

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CpG islands

Alternative DNA sites near genes; high in CG content, often nucleosome-free.

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Divergent transcription

RNA synthesis from a promoter producing antisense RNA; short, unstable.

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General transcription factors

Proteins needed for most RNA Polymerase II transcription.

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TFIID

General transcription factor with TBP and TAFs; positions RNA Polymerase II.

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TBP (TATA binding protein)

First protein to bind to the TATA box.

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Preinitiation complex

General transcription factors along with RNA Polymerase II bound to a promoter

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RNA Polymerase II

Enzyme that synthesizes mRNA in eukaryotic cells.

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TFIIA

A protein that binds close to TBP and upstream of TATA box in the pre-initiation complex.

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TFIIB

A protein involved in binding to the pre-initiation complex and positioning RNA polymerase II at transcription initiation.

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TFIIF

A protein that bind to the RNA Polymerase II.

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TFIIE

A protein that binds to the pre-initiation complex and creates a site for TFIIH.

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TFIIH

A protein that helps in forming open complex; uses ATP to melt DNA; phosphorylates Pol II.

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Transcription of 18S rRNA gene

Process of creating 18S rRNA from DNA template.

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Transcription of actin mRNA

Production of actin mRNA from DNA.

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Centromere

Structure that connects sister chromatids during cell division.

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RNA Polymerase I

Enzyme that transcribes rRNA genes.

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RNA Polymerase II

Enzyme that transcribes protein-coding genes.

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RNA Polymerase III

Enzyme that transcribes tRNA and other small RNA genes.

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Melting Temperature (Tm) of DNA

Temperature at which half the DNA strands separate.

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GC content

Percentage of guanine and cytosine bases in a DNA molecule.

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Ion concentration

Amount of ions in DNA solution affecting Tm.

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Chemicals destabilizing H-bonds

Chemicals like urea and formamide weakening DNA bonds.

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Two amino acids

Connected by a peptide bond.

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Guanine and cytosine

Connected by hydrogen bonds in DNA.

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Two glucose molecules

Connected by a glycosidic bond.

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Nucleic acids

DNA and RNA, connected by phosphodiester bonds.

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DNA Replication Fork

The point where DNA is unwound during replication.

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Study Notes

RNA Types and Functions

  • RNA is a type of nucleic acid involved in protein synthesis.
  • RNA differs from DNA in several key aspects: single-stranded, uses ribose sugar, uracil instead of thymine.
  • Various types of RNA exist, each with specific functions.

Types of RNA

  • mRNA (messenger RNA): Carries genetic information from DNA to the ribosomes. Codes for proteins.
  • rRNA (ribosomal RNA): Forms the core of ribosomes, the complex structures where proteins are made, and catalyzes protein synthesis.
  • tRNA (transfer RNA): Essential in protein synthesis, acts as an adaptor between mRNA and amino acids.
  • snRNA (small nuclear RNA): Essential in processing pre-mRNA, vital in splicing.
  • snoRNA (small nucleolar RNA): Helps in processing and modifying rRNA.
  • miRNA (microRNA): Regulates gene expression by blocking translation and causing mRNA degradation.
  • siRNA (small interfering RNA): Silences gene expression via mRNA degradation and compact chromatin structure formation.
  • piRNA (Piwi-interacting RNA): Protects germ cells from transposable elements.
  • IncRNA (long noncoding RNA): Regulates diverse cellular processes, including X-chromosome inactivation.

Central Dogma

  • DNA serves as a blueprint, and its instructions are copied into RNA by a process called transcription.
  • The instructions in RNA then direct the assembly of amino acids to create protein in a process known as translation.
  • The process is DNA ↔ RNA ↔ protein.

Gene Definition

  • A gene is a unit of DNA that contains the instructions for the synthesis of a single polypeptide chain or functional RNA molecule (e.g., tRNA).

RNA Polymerases

  • Involved in the transcription process.
  • Eukaryotic cells have three main types of RNA polymerase.
    • Their roles in transcription are distinguished by the types of RNA they produce.
  • Different RNA polymerases produce different types of RNA molecules in eukaryotic cells.

Transcription

  • RNA polymerase moves along the DNA template strand in a 3' to 5' direction.
  • The process generates a new RNA strand that is complementary to the template strand in a 5' to 3' direction. (transcription bubble is 1000-2000 nucleotides).
  • 5' capping and 3' cleavage and polyadenylation are post-transcriptional processes that modify mRNA in eukaryotic cells.

RNA Processing

  • 5' capping: Protective structure added to the 5' end of eukaryotic mRNA.
  • 3' cleavage and polyadenylation: Removes the end and adds a poly(A) tail by adding multiple adenine nucleotides to the 3' end of eukaryotic mRNA.
  • RNA splicing: Removes non-coding introns and joins coding exons together. Splicing is tightly controlled through a complex called a spliceosome.

Alternative Splicing

  • Different mRNAs may be produced by alternate arrangement of exons during RNA splicing.
  • Exons are spliced together in a variety of ways.
  • mRNA diversity is an important process for building a wider array of proteins from one base sequence.

Protein Complexes Associated with Pre-mRNAs

  • Heterogeneous ribonucleoprotein particles (hnRNPs) are protein complexes containing hnRNA and involved in pre-mRNA processing.
  • hnRNP proteins include RNA-binding domains that affect accessibility and uniformity of pre-mRNAs, participate in RNA splicing or cleavage/polyadenylation, and export from the nucleus to the cytoplasm.

Transcription Control Regions

  • Includes promoters, proximal elements, distant enhancers, and silencers.
  • They can stimulate transcription even when inverted, meaning they can be turned “on” or “off”.
  • These types of control regions are often cell-type specific making sure proteins build at needed times and places in different cells.

Promoters and Initiation Sites

  • Promoter sequences are located upstream of the initiation site.
  • The TATA box and initiators regulate start site selection.
  • CpG islands are often found in the promoters of mammalian genes where numerous CG sequences appear together.
  • CpG islands are unique to certain areas of the genome indicating potential initiation regions.
  • Divergent transcription can start in opposite directions from these islands.

General Transcription Factors

  • Several proteins are required to initiate transcription by RNA polymerase II. (example: TBP, TAFs)
  • These factors position RNA polymerase II to the start site leading to initiation.

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