Transcription

PROTEIN SYNTHESIS
Francis Ian L. Salaver, RMT BMLS 3A

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Protein Synthesis (Gene Expression) Notes

Proteins
Proteins make up all living materials
Proteins are composed of amino acids there are 20 different amino acids Different proteins are made by combining these 20 amino acids in different combinations
DNA and Genes

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Gene
DNA
Control Of Proteins
Protein
Trait
Proteins
Cys Pro Glu A M K
Cys
Glu
His
Met
Phe
His
DNA
DNA contains genes, sequences of nucleotide bases These Genes code for polypeptides (proteins) Proteins are used to build cells and do much of the work inside cells
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Genes & Proteins

Proteins are made of amino acids linked together by peptide bonds 20 different amino acids exist
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Amino Acid Structure
Polypeptides
Amino acid chains are called polypeptides
Glu
Cys Pro
Glu
M K
Cys
His
Met
Phe
His
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Fact!
All living things synthesize proteins. In fact, the types of proteins that a cell synthesizes determine the kind of cell it is.
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DNA Begins the Process

DNA is found inside the nucleus Proteins, however, are made in the cytoplasm of cells by organelles called ribosomes Ribosomes may be free in the cytosol or attached to the surface of rough ER
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DNA Transcription
Proteins are manufactured (made) by the ribosomes
Starting with DNA

DNA s code must be copied and taken to the cytosol In the cytoplasm, this code must be read so amino acids can be assembled to make polypeptides (proteins) This process is called PROTEIN SYNTHESIS
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Roles of RNA and DNA

DNA is the MASTER PLAN RNA is the BLUEPRINT of the Master Plan
CENTRAL DOGMA OF LIFE

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CENTRAL DOGMA OF LIFE

Transcription = DNA to mRNA
Translation = mRNA to Proteins
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Expression of Genes
Some genes are transcribed in large quantities because we need large amount of this protein Some genes are transcribed in small quantities because we need only a small amount of this protein
RNA Differs from DNA

RNA has a sugar ribose DNA has a sugar deoxyribose
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Other Differences
RNA contains the base uracil (U) DNA has thymine (T) RNA molecule is single-stranded DNA is doublestranded
DNA
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Structure of RNA
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RNA Differences
DNA-RNA Complementarity
First evidence, it was shown that the RNAs produced by various organisms have base ratios very similar to the base ratios in the same organismsDNA
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DNA-RNA Complementarity
Second line of evidence comes from experiments by B. Hall, S. Spiegelman, and others using DNA-RNA hybridization.
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In another experiment, DNA-RNA hybridization showed that bacteriophage infection led to the production of phagespecific messenger RNA. Gene-sized pieces of RNA extracted from Escherichia coli before and after bacteriophage T2 infection were tested to see if they hybridized with the DNA of the T2 phage or with the DNA of the E. coli cell. The RNA in the E. coli cell was found to hybridize with the E. coli DNA before infection but with the T2 DNA after infection. Thus it is apparent that when the phage attacks the E. coli cell, it starts to manufacture RNA complementary to its own DNA and stops the E. coli DNA from serving as a template.
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Three Types of RNA

Messenger RNA (mRNA) copies DNAs code & carries the genetic information to the ribosomes
Ribosomal RNA (rRNA), along with protein, makes up the ribosomes

Transfer RNA (tRNA) transfers amino acids to the ribosomes where proteins are synthesized 31
Ribosomal RNA (rRNA)

rRNA is a single strand 100 to 3000 nucleotides long Globular in shape Made inside the nucleus of a cell Associates with proteins to form ribosomes Site of protein Synthesis
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Messenger RNA (mRNA)

Carries the information for a specific protein Made up of 500 to 1000 nucleotides long Sequence of 3 bases called codon AUG methionine or start codon UAA, UAG, or UGA stop codons
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Transfer RNA (tRNA)

Clover-leaf shape Single stranded molecule with attachment site at one end for an amino acid Opposite end has three nucleotide bases called the anticodon
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Codons and Anticodons

The 3 bases of an anticodon are complementary to the 3 bases of a codon Example: Codon ACU Anticodon UGA
UGA
ACU
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Transcription
the process of synthesizing RNA from a DNA template using the rules of complementarity
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Transcription versus DNA Replication

DNA polymerases and some of the other proteins involved in DNA replication bind to DNA, but they do not necessarily bind to any specific sequences. Proteins that recognize specific DNA sequences are critically important to the transcriptional process.
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Transcription vs Replication
Open and unwind a portion of the DNA 1 strand of the DNA acts as a template
Differences
RNA strand does not stay paired with DNA
DNA re-coils and RNA is single stranded
RNA is shorter than DNA
Part of DNA temporarily unzips and is used as a template to assemble complementary nucleotides into messenger RNA (mRNA).
Protein synthesis (Prokaryotes)

RNA Polymerase
Uncoils the DNA (bacteria) Core component and sigma factor Catalyzes the formation of the phosphodiester bonds between the nucleotides (sugar to phosphate)
RNA Synthesis
RNA pol opens the DNA double helix and creates the template RNA pol moves nt by nt, unwinds the DNA as it goes Will stop when it encounters a STOP codon, RNA pol leaves, releasing the RNA strand
Sigma () Factor
Part of the bacterial RNA polymerase that helps it recognize the promoter Released after about 10 nucleotides of RNA are linked together Rejoins with a released RNA polymerase to look for a new promoter
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NOTE!
RNA polymerase must be able to recognize both the beginnings and the ends of genes (or gene groups) on the DNA double helix in order to initiate and terminate transcription. It must also be able to recognize the correct DNA strand to avoid transcribing the DNA strand that is not informational. RNA polymerase accomplishes those tasks by recognizing certain start and stop signals in DNA, called initiation and termination sequences, respectively
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Promoter
The DNA region that RNA polymerase associates with immediately before beginning transcription is known as the promoter. The promoter is an important part of gene expression in both prokaryotes and eukaryotes. Promoters contain the information for transcription initiation and are the major sites in which gene expression is controlled.
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Termination Sequence
Termination of transcription comes about when the polymerase enzyme recognizes a DNA region known as a terminator sequence
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Fact
The RNA polymerase molecule covers a region of about sixty base pairs of DNA. This was determined by causing the polymerase to bind to DNA and then digesting the mixture with nucleases, in a technique known as footprinting. Conserved versus Consensus sequence
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Termination
The polymerase moves down the DNA until the RNA polymerase reaches a stop signal, or terminator sequence. Two types of terminators, rho-dependent and rhoindependent, differ in their dependency on the rho protein.
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Eukaryotic Transcription
Transcription occurs in the nucleus in eukaryotes,
How does the process of transcription begin?

The DNA serves as the template for producing an RNA transcript or copy of information stored on the DNA molecule.
The DNA molecule must open up and allow an enzyme called RNA polymerase read and connect together the sequence of nucleotides in the proper order.

After the RNA transcript is made, it is not yet ready to leave the nucleus. Regions called introns (noncoding) are snipped out with enzymes and degraded, leaving behind exons (coding regions of RNA) which are spliced together.
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A sequence of nucleotides (AAAA to 3 end of RNA transcript are added, polyadenylation, which allows the product to leave the nucleus Mature RNA transcript. If the mature RNA transcript carries the message for producing proteins, then it becomes mRNA.
RNA Processing
Eukaryotic cells process the RNA in the nucleus before it is moved to the cytoplasm for protein synthesis The RNA that is the direct copy of the DNA is the primary transcript 2 methods used to process primary transcripts to increase the stability of mRNA being exported to the cytoplasm
RNA capping Polyadenylation
RNA Processing
RNA capping happens at the 5 end of the RNA, usually adds a methylgaunosine shortly after RNA polymerase makes the 5 end of the primary transcript Polyadenylation modifies the 3 end of the primary transcript by the addition of a string of As
At the 5' end, a cap is added consisting of a modified GTP (guanosine triphosphate). This occurs at the beginning of transcription. The 5' cap is used as a recognition signal for ribosomes to bind to the mRNA. At the 3' end, a poly(A) tail of 150 or more adenine nucleotides is added. The tail plays a role in the stability of the mRNA.
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mRNA then goes through the pores of the nucleus with the DNA code and attaches to the ribosome.
Review~~
One student will be called to explain following slides.
One student per slide
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Pathway to Making a Protein

DNA
mRNA tRNA (ribosomes) Protein
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Protein Synthesis
The production or
synthesis of polypeptide chains (proteins) Two phases: Transcription & Translation mRNA must be processed before it leaves the nucleus of eukaryotic cells
Transcription
The process of copying the sequence of one strand of DNA, the template strand mRNA copies the template strand Requires the enzyme RNA Polymerase
Template Strand
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Transcription
During transcription, RNA polymerase binds to DNA and separates the DNA strands RNA Polymerase then uses one strand of DNA as a template to assemble nucleotides into RNA
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Transcription
Promoters are regions on DNA that show where RNA Polymerase must bind to begin the Transcription of RNA Called the TATA box Specific base sequences act as signals to stop Called the cmassengale copyright termination signal
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RNA Polymerase
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mRNA Processing
After the DNA is transcribed into RNA, editing must be done to the nucleotide chain to make the RNA functional Introns, non-functional segments of DNA are snipped out of the chain
mRNA Editing
Exons, segments of DNA that code for proteins, are then rejoined by the enzyme ligase A guanine triphosphate cap is added to the 5 end of the newly copied mRNA A poly A tail is added to the 3 end of the RNA The newly processed mRNA can then leave the nucleus copyright cmassengale 76
Result of Transcription
CAP
New Transcript
Tail
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Messenger RNA (mRNA)

start codon mRNA
A U G G G C U C C A U C G G C G C A U A A
codon 1 codon 2
glycine
codon 3
serine
codon 4
isoleucine
codon 5
glycine
codon 6
alanine
codon 7
stop codon
protein methionine
Primary structure of a protein

aa1 aa2 aa3 aa4 aa5 aa6
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PROTEIN SYNTHESIS

Francis Ian L. Salaver, RMT BMLS 3A

Protein Synthesis (Gene Expression) Notes

DNA and Genes

Genes & Proteins

Amino Acid Structure

DNA Begins the Process

Proteins are manufactured (made) by the ribosomes

Starting with DNA

Roles of RNA and DNA

CENTRAL DOGMA OF LIFE

CENTRAL DOGMA OF LIFE

RNA Differs from DNA

Three Types of RNA

Ribosomal RNA (rRNA), along with protein, makes up the ribosomes

Ribosomal RNA (rRNA)

Messenger RNA (mRNA)

Transfer RNA (tRNA)

Codons and Anticodons

Transcription versus DNA Replication

RNA is shorter than DNA

Protein synthesis (Prokaryotes)

Transcription occurs in the nucleus in eukaryotes,

How does the process of transcription begin?

How does the process of transcription begin?

How does the process of transcription begin?

Pathway to Making a Protein

Messenger RNA (mRNA)

Primary structure of a protein

peptidecopyright cmassengale bonds

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