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DNA/RNA…… part 3

                                                            By


                                       Taghrid El Abaseri MD, PhD.

                                                 April 2nd 2012
                                             SCU Dentistry School D1




                                                                                               1




                                                                                                   1


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The Replication Fork

                    Only one strand of DNA is synthesized in a continuous manner in the
                       direction of overall DNA replication 5 to 3
                    The leading strand is the strand of DNA that is synthesized continuously in
                       the direction of movement of the replication fork.
                    Okazaki fragments are small pieces of newly synthesized DNA that are joined
                       to form an intact new DNA strand.
                    DNA ligase is an enzyme that seals breaks in DNA strands.
                    The lagging strand is the strand of DNA synthesized opposite to the direction
                       of movement of the replication fork, by ligation of Okazaki fragments.
                    Short fragments of RNA serve as primers for DNA replication.
                    Primase is an enzyme that synthesizes short fragments of RNA
                       complementary to the lagging strand template at the replication fork.
                    RNase H is an enzyme that degrades the RNA strand of RNA-DNA hybrids,
                      and 5 to 3 exonucleases.




                                                                                                    2




                                                                                                        2


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Synthesis of leading and lagging strands of DNA




                                                                                               3




                                                                                                   3


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Initiation of Okazaki fragments with RNA primers




                                                                                               4




                                                                                                   4


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Removal of RNA primers and joining of Okazaki fragments




                                                                                               5




                                                                                                   5


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Expression of Genetic Information

                    Genes act by determining the structure of proteins, which are responsible for
                       directing cell metabolism through their activity as enzymes.

                    Proteins, in turn, are polymers of 20 amino acids, the sequence of which
                       determines their structure and function.

                    The first direct link between a genetic mutation and an alteration in the amino
                       acid sequence of a protein was made in 1957.
                         •   Individuals with the inherited disease sickle-cell anemia have
                             hemoglobin molecules that differ from normal ones by a single amino
                             acid substitution.




                                                                                                      6




                                                                                                          6


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Colinearity of Genes and Proteins

                    The simplest hypothesis to account for the relationship between genes and
                       enzymes was that the order of nucleotides in DNA specified the order of
                       amino acids in a protein.

                    Another hypothesis predicted that different mutations within a single gene
                       could alter different amino acids in the encoded protein.




                                                                                                 7




                                                                                                     7


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Synthesis of RNA from DNA

                    Although the sequence of nucleotides in DNA
                        appeared to specify the order of amino
                        acids in proteins, it did not necessarily
                        follow that DNA itself directs protein
                        synthesis.
                    RNA appeared a likely candidate for such an
                        intermediate because the similarity of its
                        structure to that of DNA suggested that
                        RNA could be synthesized from a DNA
                        template.


                    RNA polymerase is the principal
                      enzyme responsible for RNA
                      synthesis.




                                                                                               8




                                                                                                   8


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Eukaryotic RNA Polymerases and General Transcription Factors


                    eukaryotic cells contain multiple different RNA polymerases that transcribe
                      distinct classes of genes.

                    Transcription in eukaryotes takes place on chromatin rather than on free DNA.

                    Regulation of chromatin structure is an important factor in the transcriptional
                      activity of eukaryotic genes.

                    Eukaryotic cells contain three distinct nuclear RNA polymerases that
                      transcribe different classes of genes.
                    Although all three of the nuclear RNA polymerases recognize different
                      promoters and transcribe distinct classes of genes, they share several
                      features in common with each other




                                                                                                      9


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10


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Transcription in eukaryotes

                    A promoter is the DNA sequence to which RNA polymerase binds to initiate
                      transcription of a gene.

                    After transcription stops, the RNA is released from the polymerase, and the
                      enzyme dissociates from its DNA template.




                                                                                                  11


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Transcription by RNA polymerase




                                                                                               12


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Processing of mRNA in Eukaryotes


                    Pre-mRNA is the primary transcript that is processed to form messenger RNA
                      in eukaryotic cells.

                    A 7-methylguanosine cap is what is added during the modification of the 5
                      end of a transcript.

                    A poly-A tail is a tract of about 200 adenine nucleotides added to the 3 ends
                      of eukaryotic mRNAs.

                    Polyadenylation is the process of adding a poly-A tail to a pre-mRNA.




                                                                                                     13


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Processing of eukaryotic messenger RNAs




                                                                                               14


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The Role of Messenger RNA


                    The central dogma of molecular biology states that RNA molecules are
                       synthesized from DNA templates, and proteins are synthesized from RNA
                       templates.
                         •   Transcription is the synthesis of an RNA molecule from a DNA
                             template.
                         •   Translation is the synthesis of a polypeptide chain from an mRNA
                             template.

                    Messenger RNAs (mRNAs) are RNA molecules that serve as templates for
                       protein synthesis.

                    RNA polymerase is an enzyme that catalyzes the synthesis of RNA.

                    Ribosomal RNA (rRNA) is a component of ribosomes.

                    Transfer RNAs (tRNAs) serve as adaptor molecules that align amino acids
                       along the mRNA template.




                                                                                                15




                                                                                                     15


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The Genetic Code

                    Genetic information is
                      transferred between
                      nucleic acids and
                      proteins.
                    The genetic code is the
                      correspondence that
                      takes place between
                      nucleotide triplets and
                      amino acids in
                      proteins.
                    Codons are the basic
                      units of the genetic
                      code.




                                                       Function of transfer RNA 16




                                                                                               16


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17




                                                                                                    17


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Translation of mRNA


                    Proteins are synthesized from mRNA templates by a process that has been
                       highly conserved throughout evolution.

                    Translation is carried out on ribosomes, with tRNAs serving as adaptors
                       between the mRNA template and the amino acids being incorporated into
                       protein.




                                                                                               18




                                                                                                    18


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Structure of tRNAs


                 Transfer RNAs, or tRNAs, possess
                    unique identifying sequences
                    that allow the correct amino
                    acid to be attached and aligned
                    with the appropriate codon in
                    mRNA.
                 Transfer RNAs are approximately
                    70 to 80 nucleotides long and
                    have characteristic cloverleaf
                    structures.




                                                                                               19




                                                                                                    19


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Transfer RNAs

                    Aminoacyl tRNA synthetases are a group of
                     enzymes that recognize a single amino acid, as
                     well as the correct tRNA (or tRNAs) to which that
                     amino acid should be attached.
                    After being attached to tRNA, an amino acid is
                      aligned on the mRNA template by complementary
                      base pairing between the mRNA codon and the
                      anticodon of the tRNA.




                                                                                               20




                                                                                                    20


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Ribosome structure

                    Like tRNAs, rRNAs form characteristic secondary structures by
                       complementary base pairing.
                    The role of rRNA in the formation of peptide bonds extends the catalytic
                       activities of RNA beyond self-replication to direct involvement in protein
                       synthesis.


               Ribosomes are the sites of protein
                   synthesis in both prokaryotic and
                   eukaryotic cells.

               Ribosomal RNAs, or rRNAs, are the
                   ribosomal components of ribosomes.

               The general structures of prokaryotic and
                   eukaryotic ribosomes are similar,
                   although they differ in some details.




                                                                                                    21




                                                                                                         21


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The Organization of mRNAs and the Initiation of Translation




                               The signals that identify initiation codons in eukaryotic cells.




                                                                                                  22




                                                                                                       22


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Overview of translation

                    Translation is generally divided into three stages:
                      initiation, elongation, and termination.




                                                                                               23




                                                                                                    23


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Initiation of translation in eukaryotic cells




                                                                                               24




                                                                                                    24


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Elongation stage of translation



                After the initiation complex has
                    formed, translation proceeds by
                    elongation of the polypeptide
                    chain.
                Elongation factors, which are
                    complexed to GTPs, escort the
                    aminoacyl tRNA to the ribosome.
                The next step in elongation is
                    translocation, which requires
                    another elongation factor and is
                    coupled to GTP hydrolysis.




                                                                                               25




                                                                                                    25


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Termination of translation




                 Release factors are proteins that
                 recognize stop codons and terminate
                 translation of mRNA.




                                                                                               26




                                                                                                    26


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Dna and rna_molecular_biology_part3_dent_april_2nd_2012

  • 1. DNA/RNA…… part 3 By Taghrid El Abaseri MD, PhD. April 2nd 2012 SCU Dentistry School D1 1 1 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 2. The Replication Fork Only one strand of DNA is synthesized in a continuous manner in the direction of overall DNA replication 5 to 3 The leading strand is the strand of DNA that is synthesized continuously in the direction of movement of the replication fork. Okazaki fragments are small pieces of newly synthesized DNA that are joined to form an intact new DNA strand. DNA ligase is an enzyme that seals breaks in DNA strands. The lagging strand is the strand of DNA synthesized opposite to the direction of movement of the replication fork, by ligation of Okazaki fragments. Short fragments of RNA serve as primers for DNA replication. Primase is an enzyme that synthesizes short fragments of RNA complementary to the lagging strand template at the replication fork. RNase H is an enzyme that degrades the RNA strand of RNA-DNA hybrids, and 5 to 3 exonucleases. 2 2 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 3. Synthesis of leading and lagging strands of DNA 3 3 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 4. Initiation of Okazaki fragments with RNA primers 4 4 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 5. Removal of RNA primers and joining of Okazaki fragments 5 5 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 6. Expression of Genetic Information Genes act by determining the structure of proteins, which are responsible for directing cell metabolism through their activity as enzymes. Proteins, in turn, are polymers of 20 amino acids, the sequence of which determines their structure and function. The first direct link between a genetic mutation and an alteration in the amino acid sequence of a protein was made in 1957. • Individuals with the inherited disease sickle-cell anemia have hemoglobin molecules that differ from normal ones by a single amino acid substitution. 6 6 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 7. Colinearity of Genes and Proteins The simplest hypothesis to account for the relationship between genes and enzymes was that the order of nucleotides in DNA specified the order of amino acids in a protein. Another hypothesis predicted that different mutations within a single gene could alter different amino acids in the encoded protein. 7 7 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 8. Synthesis of RNA from DNA Although the sequence of nucleotides in DNA appeared to specify the order of amino acids in proteins, it did not necessarily follow that DNA itself directs protein synthesis. RNA appeared a likely candidate for such an intermediate because the similarity of its structure to that of DNA suggested that RNA could be synthesized from a DNA template. RNA polymerase is the principal enzyme responsible for RNA synthesis. 8 8 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 9. Eukaryotic RNA Polymerases and General Transcription Factors eukaryotic cells contain multiple different RNA polymerases that transcribe distinct classes of genes. Transcription in eukaryotes takes place on chromatin rather than on free DNA. Regulation of chromatin structure is an important factor in the transcriptional activity of eukaryotic genes. Eukaryotic cells contain three distinct nuclear RNA polymerases that transcribe different classes of genes. Although all three of the nuclear RNA polymerases recognize different promoters and transcribe distinct classes of genes, they share several features in common with each other 9 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 10. 10 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 11. Transcription in eukaryotes A promoter is the DNA sequence to which RNA polymerase binds to initiate transcription of a gene. After transcription stops, the RNA is released from the polymerase, and the enzyme dissociates from its DNA template. 11 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 12. Transcription by RNA polymerase 12 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 13. Processing of mRNA in Eukaryotes Pre-mRNA is the primary transcript that is processed to form messenger RNA in eukaryotic cells. A 7-methylguanosine cap is what is added during the modification of the 5 end of a transcript. A poly-A tail is a tract of about 200 adenine nucleotides added to the 3 ends of eukaryotic mRNAs. Polyadenylation is the process of adding a poly-A tail to a pre-mRNA. 13 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 14. Processing of eukaryotic messenger RNAs 14 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 15. The Role of Messenger RNA The central dogma of molecular biology states that RNA molecules are synthesized from DNA templates, and proteins are synthesized from RNA templates. • Transcription is the synthesis of an RNA molecule from a DNA template. • Translation is the synthesis of a polypeptide chain from an mRNA template. Messenger RNAs (mRNAs) are RNA molecules that serve as templates for protein synthesis. RNA polymerase is an enzyme that catalyzes the synthesis of RNA. Ribosomal RNA (rRNA) is a component of ribosomes. Transfer RNAs (tRNAs) serve as adaptor molecules that align amino acids along the mRNA template. 15 15 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 16. The Genetic Code Genetic information is transferred between nucleic acids and proteins. The genetic code is the correspondence that takes place between nucleotide triplets and amino acids in proteins. Codons are the basic units of the genetic code. Function of transfer RNA 16 16 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 17. 17 17 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 18. Translation of mRNA Proteins are synthesized from mRNA templates by a process that has been highly conserved throughout evolution. Translation is carried out on ribosomes, with tRNAs serving as adaptors between the mRNA template and the amino acids being incorporated into protein. 18 18 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 19. Structure of tRNAs Transfer RNAs, or tRNAs, possess unique identifying sequences that allow the correct amino acid to be attached and aligned with the appropriate codon in mRNA. Transfer RNAs are approximately 70 to 80 nucleotides long and have characteristic cloverleaf structures. 19 19 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 20. Transfer RNAs Aminoacyl tRNA synthetases are a group of enzymes that recognize a single amino acid, as well as the correct tRNA (or tRNAs) to which that amino acid should be attached. After being attached to tRNA, an amino acid is aligned on the mRNA template by complementary base pairing between the mRNA codon and the anticodon of the tRNA. 20 20 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 21. Ribosome structure Like tRNAs, rRNAs form characteristic secondary structures by complementary base pairing. The role of rRNA in the formation of peptide bonds extends the catalytic activities of RNA beyond self-replication to direct involvement in protein synthesis. Ribosomes are the sites of protein synthesis in both prokaryotic and eukaryotic cells. Ribosomal RNAs, or rRNAs, are the ribosomal components of ribosomes. The general structures of prokaryotic and eukaryotic ribosomes are similar, although they differ in some details. 21 21 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 22. The Organization of mRNAs and the Initiation of Translation The signals that identify initiation codons in eukaryotic cells. 22 22 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 23. Overview of translation Translation is generally divided into three stages: initiation, elongation, and termination. 23 23 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 24. Initiation of translation in eukaryotic cells 24 24 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 25. Elongation stage of translation After the initiation complex has formed, translation proceeds by elongation of the polypeptide chain. Elongation factors, which are complexed to GTPs, escort the aminoacyl tRNA to the ribosome. The next step in elongation is translocation, which requires another elongation factor and is coupled to GTP hydrolysis. 25 25 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)
  • 26. Termination of translation Release factors are proteins that recognize stop codons and terminate translation of mRNA. 26 26 Create PDF files without this message by purchasing novaPDF printer (http://www.novapdf.com)