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Biochemistry for Medics
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                Biochemistry For Medics   1
RNA (Ribonucleic acid )

RNA is a
polymer of
ribonucleotides
linked together
by 3’-5’
phosphodiester
linkage


                  Biochemistry For Medics   2
RNA V/S DNA




              Biochemistry For Medics   3
Differences between RNA and DNA
 S.No.   RNA                                      DNA

 1)      Single stranded mainly except            Double stranded (Except for
         when self complementary                  certain viral DNA s which are
         sequences are there it forms a           single stranded)
         double stranded structure (Hair
         pin structure)
 2)      Ribose is the main sugar                 The sugar moiety is deoxy
                                                  ribose
 3)      Pyrimidine components differ.            Thymine is always there but
         Thymine is never found(Except            uracil is never found
         tRNA)
 4)      Being single stranded structure-         It does follow Chargaff's rule.
         It does not follow Chargaff’s rule       The total purine content in a
                                                  double stranded DNA is always
                                                  equal to pyrimidine content.

                             Biochemistry For Medics                                4
Differences between RNA and DNA
 S.No.   RNA                                      DNA
 5)      RNA can be easily destroyed by           DNA resists alkali action due to
         alkalies to cyclic diesters of           the absence of OH group at 2’
         mono nucleotides.                        position
 6)      RNA is a relatively a labile             DNA is a stable molecule. The
         molecule, undergoes easy and             spontaneous degradation is
         spontaneous degradation                  very 2 slow. The genetic
                                                  information can be stored for
                                                  years together without any
                                                  change.
 7)      Mainly cytoplasmic, but also             Mainly found in nucleus, extra
         present in nucleus (primary              nuclear DNA is found in
         transcript and small nuclear             mitochondria, and plasmids
         RNA)                                     etc
 8)      The base content varies from             Millions of base pairs are there
         100- 5000. The size is variable.         depending upon the organism

                               Biochemistry For Medics                               5
Differences between RNA and DNA
 S.No.   RNA                                     DNA
 9)      There are various types of RNA –        DNA is always of one type and
         mRNA, r RNA, t RNA, Sn RNA, Si          performs the function of
         RNA, mi RNA and hn RNA. These           storage and transfer of genetic
         RNAs perform different and              information.
         specific functions.
 10)     No variable physiological forms         There are variable forms of
         of RNA are found. The different         DNA (A to E and Z)
         types of RNA do not change
         their forms
 11)     RNA is synthesized from DNA, it         DNA can form DNA by
         can not form DNA(except by the          replication, it can also form
         action of reverse transcriptase).       RNA by transcription.
         It can not duplicate (except in
         certain viruses where it is a
         genomic material )
 12)     Many copies of RNA are present          Single copy of DNA is present
         per cell                                per cell.
                               Biochemistry For Medics                             6
Types of RNA
In all prokaryotic and eukaryotic
organisms, three main classes of RNA
molecules exist-
1) Messenger RNA(m RNA)
2) Transfer RNA (t RNA)
3) Ribosomal RNA (r RNA)
The other are –
o small nuclear RNA (SnRNA),
o micro RNA(mi RNA) and
o small interfering RNA(Si RNA) and
o heterogeneous nuclear RNA (hnRNA).

                 Biochemistry For Medics   7
Messenger RNA (m-RNA)




Comprises only 5% of the RNA in the cell
Most heterogeneous in size and base sequence
All members of the class function as
messengers carrying the information in a gene to
the protein synthesizing machinery
                    Biochemistry For Medics        8
Structural Characteristics of
m-RNA
 The 5’ terminal end is capped by 7-
 methyl guanosine triphosphate cap.
 The cap is involved in the
 recognition of mRNA by the
 translating machinery
 It stabilizes m RNA by protecting it
 from 5’ exonuclease

                  Biochemistry For Medics   9
Structural Characteristics of
m-RNA(contd.)
 The 3’end of most m-RNAs have a polymer of
Adenylate residues( 20-250)
 The tail prevents the attack by 3’ exonucleases
Histones and interferons do not contain poly A
tails
 On both 5’ and 3’ end there are non coding
sequences which are not translated (NCS)
The intervening region between non coding
sequences present between 5’ and 3’ end is
called coding region. This region encodes for the
synthesis of a protein.

                     Biochemistry For Medics        10
Structural Characteristics of
m-RNA




5’ cap and 3’ tail impart stability to m RNA by protecting from
specific exo nucleases.
                           Biochemistry For Medics                11
Structural Characteristics of
m-RNA(Contd.)
The m- RNA molecules are formed with the help of
DNA template during the process of transcription.
The sequence of nucleotides in m RNA is
complementary to the sequence of nucleotides on
template DNA.
The sequence carried on m -RNA is read in the form
of codons.
A codon is made up of 3 nucleotides
The m-RNA is formed after processing of
heterogeneous nuclear RNA



                     Biochemistry For Medics          12
Heterogeneous nuclear RNA
(hnRNA)
 In mammalian nuclei , hnRNA is the
immediate product of gene transcription
 The nuclear product is heterogeneous in size
(Variable) and is very large.
Molecular weight may be more than 107, while
the molecular weight of m RNA is less than 2x 106
 75 % of hnRNA is degraded in the nucleus,
only 25% is processed to mature
m RNA


                     Biochemistry For Medics        13
Heterogeneous nuclear RNA
(hnRNA)




Mature m –RNA is formed from primary transcript by
 capping, tailing, splicing and base modification.
                    Biochemistry For Medics          14
Transfer RNA (t- RNA)
Transfer RNA are the smallest of three major species of
RNA molecules
They have 74-95 nucleotide residues
 They are synthesized by the nuclear processing of a
precursor molecule
They transfer the amino acids from cytoplasm to the
protein synthesizing machinery, hence the name t RNA.
They are easily soluble , hence called “Soluble RNA or s
RNA
They are also called Adapter molecules, since they act as
adapters for the translation of the sequence of nucleotides
of the m RNA in to specific amino acids
There are at least 20 species of t RNA one corresponding
to each of the 20 amino acids required for protein
synthesis.

                           Biochemistry For Medics            15
Structural characteristics of t- RNA
 1) Primary structure- The nucleotide
 sequence of all the t RNA molecules allows
 extensive intrastand complimentarity that
 generates a secondary structure.
 2) Secondary structure- Each single t- RNA
 shows extensive internal base pairing and
 acquires a clover leaf like structure. The
 structure is stabilized by hydrogen bonding
 between the bases and is a consistent
 feature.
                   Biochemistry For Medics     16
Structural characteristics of t-
RNA
Secondary structure (Clover leaf structure)
All t-RNA contain 5 main arms or loops
which are as follows-
a) Acceptor arm
b) Anticodon arm
c) D HU arm
d) TΨ C arm
e) Extra arm
                   Biochemistry For Medics    17
Secondary structure of t- RNA
a)Acceptor arm
 The acceptor arm is at 3’ end
 It has 7 base pairs
 The end sequence is unpaired
  Cytosine, Cytosine-Adenine at the 3’ end
 The 3’ OH group terminal of Adenine binds with
  carboxyl group of amino acids
 The t RNA bound with amino acid is called
  Amino acyl t RNA
 CCA attachment is done post transcriptionally

                    Biochemistry For Medics        18
Secondary structure of t- RNA




The carboxyl group of amino acid is attached to 3’OH group of Adenine
nucleotide of the acceptor arm. The anticodon arm base pairs with the codon
present on the m- RNA
                              Biochemistry For Medics                    19
Secondary structure of t-
RNA(contd.)
b) Anticodon arm
 Lies at the opposite end of acceptor arm
5 base pairs long
Recognizes the triplet codon present in the m
RNA
Base sequence of anticodon arm is
complementary to the base sequence of m RNA
codon.
 Due to complimentarity it can bind specifically
with m RNA by hydrogen bonds.

                      Biochemistry For Medics       20
Secondary structure of t-
RNA(contd.)
c) DHU arm
 It has 3-4 base pairs
Serves as the recognition site for the enzyme (amino
acyl t RNA synthetase) that adds the amino acid to the
acceptor arm.
d) TΨC arm
This arm is opposite to DHU arm
Since it contains pseudo uridine that is why it is so
named
 It is involved in the binding of t RNA to the ribosomes


                       Biochemistry For Medics              21
Secondary structure of t-
RNA(contd.)
e) Extra arm or Variable arm
 About 75 % of t RNA molecules possess
a short extra arm
If about 3-5 base pairs are present the
t-RNA is said to be belonging to class 1.
Majority t -RNA belong to class 1.
 The t –RNA belonging to class 2 have
long extra arm, 13-21 base pairs in length.

                    Biochemistry For Medics   22
Tertiary structure of t- RNA

 The L shaped tertiary
structure is formed by
further folding of the clover
leaf due to hydrogen bonds
between T and D arms.
 The base paired double
helical stems get arranged in
to two double helical
columns, continuous and
perpendicular to one
another.
                  Biochemistry For Medics   23
Ribosomal RNA (rRNA)
The mammalian ribosome contains two major
nucleoprotein subunits—a larger one with a molecular
weight of 2.8 x 106 (60S) and a smaller subunit with a
molecular weight of 1.4 x 106 (40S).
The 60S subunit contains a 5S ribosomal RNA (rRNA), a
5.8S rRNA, and a 28S rRNA; there are also probably more
than 50 specific polypeptides.
The 40S subunit is smaller and contains a single 18S
rRNA and approximately 30 distinct polypeptide chains.
All of the ribosomal RNA molecules except the 5S rRNA
are processed from a single 45S precursor RNA molecule
in the nucleolus .
5S rRNA is independently transcribed.
                       Biochemistry For Medics            24
Ribosomal RNA (rRNA)




            Biochemistry For Medics   25
Ribosomal RNA (rRNA)
The functions of the ribosomal RNA
molecules in the ribosomal particle are not
fully understood, but they are necessary for
ribosomal assembly and seem to play key
roles in the binding of mRNA to ribosomes
and its translation
Recent studies suggest that an rRNA
component performs the peptidyl transferase
activity and thus is an enzyme (a ribozyme).

                   Biochemistry For Medics     26
Small RNA
Most of these molecules are
complexed with proteins to form
ribonucleoproteins and are distributed in
the nucleus, in the cytoplasm, or in both.
They range in size from 20 to 300
nucleotides and are present in 100,000–
1,000,000 copies per cell.


                  Biochemistry For Medics    27
Small Nuclear RNAs (snRNAs)

snRNAs, a subset of the small RNAs, are
significantly involved in mRNA processing and
gene regulation
Of the several snRNAs, U1, U2, U4, U5, and
U6 are involved in intron removal and the
processing of hnRNA into mRNA
The U7 snRNA is involved in production of
the correct 3' ends of histone mRNA—which
lacks a poly(A) tail.
                    Biochemistry For Medics     28
Small Nuclear RNAs (snRNAs).




Sn RNA s are involved in the process of splicing (intron removal) of primary
transcript to form mature m RNA. The Sn RNA s form complexes with proteins
to form Ribonucleoprotein particles called snRNPs

                              Biochemistry For Medics                     29
Micro RNAs, miRNAs, and Small
    Interfering RNAs, siRNAs
These two classes of RNAs represent a
subset of small RNAs; both play important
roles in gene regulation.
miRNAs and siRNAs cause inhibition of
gene expression by decreasing specific
protein production albeit apparently via
distinct mechanisms


                   Biochemistry For Medics   30
Micro RNAs (miRNAs)
miRNAs are typically 21–25 nucleotides in
length and are generated by nucleolytic
processing of the products of distinct
genes/transcription units
The small processed mature miRNAs
typically hybridize, via the formation of
imperfect RNA-RNA duplexes within the 3'-
untranslated regions of specific target
mRNAs, leading via unknown mechanisms to
translation arrest.

                  Biochemistry For Medics    31
Micro RNAs (miRNAs)




microRNAs, short non-coding RNAs present in all living organisms, have
been shown to regulate the expression of at least half of all human
genes. These single-stranded RNAs exert their regulatory action by
binding messenger RNAs and preventing their translation into
proteins.
                              Biochemistry For Medics                    32
Small Interfering RNAs (siRNAs)
 siRNAs are derived by the specific nucleolytic cleavage
of larger, double-stranded RNAs to again form small 21–
25 nucleotide-long products.
These short siRNAs usually form perfect RNA-RNA
hybrids with their distinct targets potentially anywhere
within the length of the mRNA where the complementary
sequence exists.
 Formation of such RNA-RNA duplexes between siRNA
and mRNA results in reduced specific protein production
because the siRNA-mRNA complexes are degraded by
dedicated nucleolytic machinery;
 some or all of this mRNA degradation occurs in specific
organelles termed P bodies.
                        Biochemistry For Medics             33
Small Interfering RNAs (siRNAs)




Small interfering RNA (siRNA) are 20-25 nucleotide-long double-stranded RNA
molecules that have a variety of roles in the cell. They are involved in the RNA
interference (RNAi) pathway, where it interferes with the expression of a
specific gene by hybridizing to its corresponding RNA sequence in the
target mRNA. This then activates the degrading mRNA. Once the target
mRNA is degraded, the mRNA cannot be translated into protein.
                                  Biochemistry For Medics                      34
Significance of mi RNAs and si
RNAs
Both miRNAs and siRNAs represent
exciting new potential targets for
therapeutic drug development in humans.
In addition, siRNAs are frequently used to
decrease or "knock-down" specific protein
levels in experimental procedures in the
laboratory, an extremely useful and
powerful alternative to gene-knockout
technology.
                  Biochemistry For Medics     35
Summary
RNA exists in several different single-stranded
structures, most of which are directly or indirectly
involved in protein synthesis or its regulation.
The linear array of nucleotides in RNA consists of A, G,
C, and U, and the sugar moiety is ribose.
The major forms of RNA include messenger RNA
(mRNA), ribosomal RNA (rRNA), transfer RNA (tRNA),
and small nuclear RNAs (snRNAs; miRNAs).
 Certain RNA molecules act as catalysts (ribozymes).
miRNAs and siRNAs represent exciting new potential
targets for therapeutic drug development in humans.



                        Biochemistry For Medics             36

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RNA- Structure, Types and Functions

  • 2. RNA (Ribonucleic acid ) RNA is a polymer of ribonucleotides linked together by 3’-5’ phosphodiester linkage Biochemistry For Medics 2
  • 3. RNA V/S DNA Biochemistry For Medics 3
  • 4. Differences between RNA and DNA S.No. RNA DNA 1) Single stranded mainly except Double stranded (Except for when self complementary certain viral DNA s which are sequences are there it forms a single stranded) double stranded structure (Hair pin structure) 2) Ribose is the main sugar The sugar moiety is deoxy ribose 3) Pyrimidine components differ. Thymine is always there but Thymine is never found(Except uracil is never found tRNA) 4) Being single stranded structure- It does follow Chargaff's rule. It does not follow Chargaff’s rule The total purine content in a double stranded DNA is always equal to pyrimidine content. Biochemistry For Medics 4
  • 5. Differences between RNA and DNA S.No. RNA DNA 5) RNA can be easily destroyed by DNA resists alkali action due to alkalies to cyclic diesters of the absence of OH group at 2’ mono nucleotides. position 6) RNA is a relatively a labile DNA is a stable molecule. The molecule, undergoes easy and spontaneous degradation is spontaneous degradation very 2 slow. The genetic information can be stored for years together without any change. 7) Mainly cytoplasmic, but also Mainly found in nucleus, extra present in nucleus (primary nuclear DNA is found in transcript and small nuclear mitochondria, and plasmids RNA) etc 8) The base content varies from Millions of base pairs are there 100- 5000. The size is variable. depending upon the organism Biochemistry For Medics 5
  • 6. Differences between RNA and DNA S.No. RNA DNA 9) There are various types of RNA – DNA is always of one type and mRNA, r RNA, t RNA, Sn RNA, Si performs the function of RNA, mi RNA and hn RNA. These storage and transfer of genetic RNAs perform different and information. specific functions. 10) No variable physiological forms There are variable forms of of RNA are found. The different DNA (A to E and Z) types of RNA do not change their forms 11) RNA is synthesized from DNA, it DNA can form DNA by can not form DNA(except by the replication, it can also form action of reverse transcriptase). RNA by transcription. It can not duplicate (except in certain viruses where it is a genomic material ) 12) Many copies of RNA are present Single copy of DNA is present per cell per cell. Biochemistry For Medics 6
  • 7. Types of RNA In all prokaryotic and eukaryotic organisms, three main classes of RNA molecules exist- 1) Messenger RNA(m RNA) 2) Transfer RNA (t RNA) 3) Ribosomal RNA (r RNA) The other are – o small nuclear RNA (SnRNA), o micro RNA(mi RNA) and o small interfering RNA(Si RNA) and o heterogeneous nuclear RNA (hnRNA). Biochemistry For Medics 7
  • 8. Messenger RNA (m-RNA) Comprises only 5% of the RNA in the cell Most heterogeneous in size and base sequence All members of the class function as messengers carrying the information in a gene to the protein synthesizing machinery Biochemistry For Medics 8
  • 9. Structural Characteristics of m-RNA The 5’ terminal end is capped by 7- methyl guanosine triphosphate cap. The cap is involved in the recognition of mRNA by the translating machinery It stabilizes m RNA by protecting it from 5’ exonuclease Biochemistry For Medics 9
  • 10. Structural Characteristics of m-RNA(contd.)  The 3’end of most m-RNAs have a polymer of Adenylate residues( 20-250)  The tail prevents the attack by 3’ exonucleases Histones and interferons do not contain poly A tails  On both 5’ and 3’ end there are non coding sequences which are not translated (NCS) The intervening region between non coding sequences present between 5’ and 3’ end is called coding region. This region encodes for the synthesis of a protein. Biochemistry For Medics 10
  • 11. Structural Characteristics of m-RNA 5’ cap and 3’ tail impart stability to m RNA by protecting from specific exo nucleases. Biochemistry For Medics 11
  • 12. Structural Characteristics of m-RNA(Contd.) The m- RNA molecules are formed with the help of DNA template during the process of transcription. The sequence of nucleotides in m RNA is complementary to the sequence of nucleotides on template DNA. The sequence carried on m -RNA is read in the form of codons. A codon is made up of 3 nucleotides The m-RNA is formed after processing of heterogeneous nuclear RNA Biochemistry For Medics 12
  • 13. Heterogeneous nuclear RNA (hnRNA)  In mammalian nuclei , hnRNA is the immediate product of gene transcription  The nuclear product is heterogeneous in size (Variable) and is very large. Molecular weight may be more than 107, while the molecular weight of m RNA is less than 2x 106  75 % of hnRNA is degraded in the nucleus, only 25% is processed to mature m RNA Biochemistry For Medics 13
  • 14. Heterogeneous nuclear RNA (hnRNA) Mature m –RNA is formed from primary transcript by capping, tailing, splicing and base modification. Biochemistry For Medics 14
  • 15. Transfer RNA (t- RNA) Transfer RNA are the smallest of three major species of RNA molecules They have 74-95 nucleotide residues  They are synthesized by the nuclear processing of a precursor molecule They transfer the amino acids from cytoplasm to the protein synthesizing machinery, hence the name t RNA. They are easily soluble , hence called “Soluble RNA or s RNA They are also called Adapter molecules, since they act as adapters for the translation of the sequence of nucleotides of the m RNA in to specific amino acids There are at least 20 species of t RNA one corresponding to each of the 20 amino acids required for protein synthesis. Biochemistry For Medics 15
  • 16. Structural characteristics of t- RNA 1) Primary structure- The nucleotide sequence of all the t RNA molecules allows extensive intrastand complimentarity that generates a secondary structure. 2) Secondary structure- Each single t- RNA shows extensive internal base pairing and acquires a clover leaf like structure. The structure is stabilized by hydrogen bonding between the bases and is a consistent feature. Biochemistry For Medics 16
  • 17. Structural characteristics of t- RNA Secondary structure (Clover leaf structure) All t-RNA contain 5 main arms or loops which are as follows- a) Acceptor arm b) Anticodon arm c) D HU arm d) TΨ C arm e) Extra arm Biochemistry For Medics 17
  • 18. Secondary structure of t- RNA a)Acceptor arm  The acceptor arm is at 3’ end  It has 7 base pairs  The end sequence is unpaired Cytosine, Cytosine-Adenine at the 3’ end  The 3’ OH group terminal of Adenine binds with carboxyl group of amino acids  The t RNA bound with amino acid is called Amino acyl t RNA  CCA attachment is done post transcriptionally Biochemistry For Medics 18
  • 19. Secondary structure of t- RNA The carboxyl group of amino acid is attached to 3’OH group of Adenine nucleotide of the acceptor arm. The anticodon arm base pairs with the codon present on the m- RNA Biochemistry For Medics 19
  • 20. Secondary structure of t- RNA(contd.) b) Anticodon arm  Lies at the opposite end of acceptor arm 5 base pairs long Recognizes the triplet codon present in the m RNA Base sequence of anticodon arm is complementary to the base sequence of m RNA codon.  Due to complimentarity it can bind specifically with m RNA by hydrogen bonds. Biochemistry For Medics 20
  • 21. Secondary structure of t- RNA(contd.) c) DHU arm  It has 3-4 base pairs Serves as the recognition site for the enzyme (amino acyl t RNA synthetase) that adds the amino acid to the acceptor arm. d) TΨC arm This arm is opposite to DHU arm Since it contains pseudo uridine that is why it is so named  It is involved in the binding of t RNA to the ribosomes Biochemistry For Medics 21
  • 22. Secondary structure of t- RNA(contd.) e) Extra arm or Variable arm  About 75 % of t RNA molecules possess a short extra arm If about 3-5 base pairs are present the t-RNA is said to be belonging to class 1. Majority t -RNA belong to class 1.  The t –RNA belonging to class 2 have long extra arm, 13-21 base pairs in length. Biochemistry For Medics 22
  • 23. Tertiary structure of t- RNA  The L shaped tertiary structure is formed by further folding of the clover leaf due to hydrogen bonds between T and D arms.  The base paired double helical stems get arranged in to two double helical columns, continuous and perpendicular to one another. Biochemistry For Medics 23
  • 24. Ribosomal RNA (rRNA) The mammalian ribosome contains two major nucleoprotein subunits—a larger one with a molecular weight of 2.8 x 106 (60S) and a smaller subunit with a molecular weight of 1.4 x 106 (40S). The 60S subunit contains a 5S ribosomal RNA (rRNA), a 5.8S rRNA, and a 28S rRNA; there are also probably more than 50 specific polypeptides. The 40S subunit is smaller and contains a single 18S rRNA and approximately 30 distinct polypeptide chains. All of the ribosomal RNA molecules except the 5S rRNA are processed from a single 45S precursor RNA molecule in the nucleolus . 5S rRNA is independently transcribed. Biochemistry For Medics 24
  • 25. Ribosomal RNA (rRNA) Biochemistry For Medics 25
  • 26. Ribosomal RNA (rRNA) The functions of the ribosomal RNA molecules in the ribosomal particle are not fully understood, but they are necessary for ribosomal assembly and seem to play key roles in the binding of mRNA to ribosomes and its translation Recent studies suggest that an rRNA component performs the peptidyl transferase activity and thus is an enzyme (a ribozyme). Biochemistry For Medics 26
  • 27. Small RNA Most of these molecules are complexed with proteins to form ribonucleoproteins and are distributed in the nucleus, in the cytoplasm, or in both. They range in size from 20 to 300 nucleotides and are present in 100,000– 1,000,000 copies per cell. Biochemistry For Medics 27
  • 28. Small Nuclear RNAs (snRNAs) snRNAs, a subset of the small RNAs, are significantly involved in mRNA processing and gene regulation Of the several snRNAs, U1, U2, U4, U5, and U6 are involved in intron removal and the processing of hnRNA into mRNA The U7 snRNA is involved in production of the correct 3' ends of histone mRNA—which lacks a poly(A) tail. Biochemistry For Medics 28
  • 29. Small Nuclear RNAs (snRNAs). Sn RNA s are involved in the process of splicing (intron removal) of primary transcript to form mature m RNA. The Sn RNA s form complexes with proteins to form Ribonucleoprotein particles called snRNPs Biochemistry For Medics 29
  • 30. Micro RNAs, miRNAs, and Small Interfering RNAs, siRNAs These two classes of RNAs represent a subset of small RNAs; both play important roles in gene regulation. miRNAs and siRNAs cause inhibition of gene expression by decreasing specific protein production albeit apparently via distinct mechanisms Biochemistry For Medics 30
  • 31. Micro RNAs (miRNAs) miRNAs are typically 21–25 nucleotides in length and are generated by nucleolytic processing of the products of distinct genes/transcription units The small processed mature miRNAs typically hybridize, via the formation of imperfect RNA-RNA duplexes within the 3'- untranslated regions of specific target mRNAs, leading via unknown mechanisms to translation arrest. Biochemistry For Medics 31
  • 32. Micro RNAs (miRNAs) microRNAs, short non-coding RNAs present in all living organisms, have been shown to regulate the expression of at least half of all human genes. These single-stranded RNAs exert their regulatory action by binding messenger RNAs and preventing their translation into proteins. Biochemistry For Medics 32
  • 33. Small Interfering RNAs (siRNAs)  siRNAs are derived by the specific nucleolytic cleavage of larger, double-stranded RNAs to again form small 21– 25 nucleotide-long products. These short siRNAs usually form perfect RNA-RNA hybrids with their distinct targets potentially anywhere within the length of the mRNA where the complementary sequence exists.  Formation of such RNA-RNA duplexes between siRNA and mRNA results in reduced specific protein production because the siRNA-mRNA complexes are degraded by dedicated nucleolytic machinery;  some or all of this mRNA degradation occurs in specific organelles termed P bodies. Biochemistry For Medics 33
  • 34. Small Interfering RNAs (siRNAs) Small interfering RNA (siRNA) are 20-25 nucleotide-long double-stranded RNA molecules that have a variety of roles in the cell. They are involved in the RNA interference (RNAi) pathway, where it interferes with the expression of a specific gene by hybridizing to its corresponding RNA sequence in the target mRNA. This then activates the degrading mRNA. Once the target mRNA is degraded, the mRNA cannot be translated into protein. Biochemistry For Medics 34
  • 35. Significance of mi RNAs and si RNAs Both miRNAs and siRNAs represent exciting new potential targets for therapeutic drug development in humans. In addition, siRNAs are frequently used to decrease or "knock-down" specific protein levels in experimental procedures in the laboratory, an extremely useful and powerful alternative to gene-knockout technology. Biochemistry For Medics 35
  • 36. Summary RNA exists in several different single-stranded structures, most of which are directly or indirectly involved in protein synthesis or its regulation. The linear array of nucleotides in RNA consists of A, G, C, and U, and the sugar moiety is ribose. The major forms of RNA include messenger RNA (mRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), and small nuclear RNAs (snRNAs; miRNAs).  Certain RNA molecules act as catalysts (ribozymes). miRNAs and siRNAs represent exciting new potential targets for therapeutic drug development in humans. Biochemistry For Medics 36