Want to Learn about Ribonucleic Acid (RNA)?



Introduction

RNA types and functions

mRNA the DNA's messenger

 mRNA goes to make a protein

Interesting facts about RNA

 RNA technologies


Introduction

Ribonucleic acid is a long polymer of nucleotides found in the nucleus but mainly in the cytoplasm of a cell; it transmits genetic information from DNA to the cytoplasm and is involved with the synthesis of proteins that control chemical processes in the cell. Do you know that DNA is like the boss of RNA. While DNA stays in it's office (Nucleus), it sends RNA to the workstations (Cytoplasm) with its messages. Because DNA does not leave the nucleus of a cell, it must have a messanger to carry the genetic code to other places in the cell; that's RNA!. 

RNA is made up of  Ribose (a pentose = sugar with 5 carbons), Phosphoric acid and Organic bases; Purines (Adenine and Guanine) and Pyrimidines (Cytosine and Uracil). RNA exists largely as single nucleotide chains in living cells. The RNA strand is made up of alternating molecules of ribose sugar and the phosphate (See figures below). The nitrogen bases are attached to the sugar molecules in the strand and ’stick out’ laterally as in DNA. A sugar, a nitrogenous base and a phosphate together form a ribonucleotide and a RNA molecule is a polymer of ribonucleotides. Although RNA exists generally as single polynucleotide chain, some segments of RNA molecules may pair temporarily in double-helical form or may fold back on themselves to set up extensive double-helical regions. These fold-back double helices and their arrangement are  important to RNA functions.

 This is a basic structure of a RNA molecule        


 structural features of RNA

Difference between DNA and RNA structures (See figures below)

 

Characters

DNA

RNA

1.

Molecule

Double stranded, helical

Single stranded, straight or variously folded and twisted.

2.

Pentose sugar

Deoxyribose

Ribose

3.

Pyrimidine base

Thymine

Uracil

4.

Complementary base pairing

Always present and exists between A = T and G = C

Normally absent, but may be present in twisted segments of a molecule.

If present, pairing is between A = U and G = C

5.

Ratio of Purines: Pyrimidines

Always 1:1

Not necessarily 1:1

Compare the strucutre of a DNA and a RNA molecule based on the information in table above



Types of RNAs and Functions

While the function of DNA is to store genetic information, different types of RNAs are involved in a series of interconnected chores to translate the information from DNA into proteins. Proteins are the ultimate controllers of cell functions, structures and characteristics e.g. your hair, eye and skin color, height etc.

There are many different types of RNAs depending on the type of a organism.

  1. messengerRNA (mRNA): Carries the genetic information out of the nucleus into cytoplasm for protein synthesis.
  2. transferRNA (tRNA): Decodes the information in mRNA.
  3. ribosomalRNA (rRNA): Makes up ribosome, constitutes 50% of a ribosome, which is a molecular assembly involved in protein synthesis.
  4. catalyticRNAs (cRNA): Catalyses many reactions in the cytoplasm of the cell.
  5. Small Nuclear RNA (snRNA): The snRNAs have various roles in the processing of the other classes of RNA.
  6. Small Nucleolar RNA (snoRNA): There are probably over 100 of them are found in the nucleolus where they are involved in several functions including making ribosomes.

The most important RNAs are mRNA, tRNA and rRNA.

 mRNA: This is called messenger RNA because it carries information for protein synthesis from the DNA to the ribosomes in the cytoplasm (the site of protein synthesis).  Just like a mailman!. mRNA constitutes about 3-5% of the total RNA. It is produced on one of the DNA strands in a process called transcription. Hence, the base sequence of mRNA is complementary to that of the DNA strand. The bases on the mRNA strand are organized into triplets. Each triplet consists of a sequence of three consecutive nitrogenous bases called a codon (code word). Each codon specifies one amino acid from wich proteins are made. The sequence of codons on the mRNA strand is called the mRNA language. It indicates the sequence of amino acids for the synthesis of a protein. The mRNA language begins with the codon AUG (initiation codon or starting codon) and ends with UAA, UAG or UGA (stop codons).


Role of mRNA in protein synthesis

 1. Represents the sequence of codons (mRNA language) from the DNA strand.

2. Brings the sequence to the ribosomes (site of protein synthesis) in the cytoplasm.

3. Provides the sequence for the synthesis of specific protein from the amino acids (found in cytoplasm).

 rRNA : I compare rRNA to a construction worker because rRNA makes up  ribosomes - The  molecular factory  involved in protein synthesis and hence is called ribosimal (r) RNA.  It is present in the cytoplasm and forms about 80% of the total RNA. The single-stranded molecule of rRNA is variously folded and twisted upon itself in certain regions forming a secondary structure. In such folded regions, complementary bases form pairs and are joined by hydrogen bonds. The secondary structure of rRNA is a complex pattern of short double-stranded stems, interspersed with unpaired single-stranded loops and bubbles (see figure below).

Structure of rRNA.

Role of rRNA in protein synthesis

The rRNA forms complex with various proteins and make a structure called ribosome, and this complex reads the coded sequence in mRNA to link amino acids together into particular protiens.

1. It provides proper binding sites for the mRNA.

2. It orients the mRNA in such a way that its nitrogen base triplets or codons are properly read or translated.

3. It also releases tRNA after the transfer of activated amino acid to the ribosome complex.

4. It protects the mRNA strand from the action of enzymes (nucleases) which can destroy it.

5. It protects the growing  polypeptide (protein) chain from proteolytic enzymes.

 tRNA:Transports amino acids floating in the cytoplasm to ribosomes for protein synthesis (So, it's function compares to a transport vehicle). tRNA consistutes about 10 to 20% of the total RNA of the cell. tRNA strand is folded upon itself forming loops. It results in either a clover leaf pattern or hair pin pattern (see figure below). One end of the strand has guanine, while the other end carries the CCA combination of nitrogen bases. A triplet of nitrogen bases called anticodon is present on one of the loops. The anticodon pairs with the complementary codon on the mRNA molecule.

structure of tRNA


Role of tRNA in protein synthesis

1. tRNA carries the required specific amino acids from cytoplasm to the ribosome (the site of protein synthesis). Each type of amino acid is carried by a specific type of tRNA.

2. In the ribosome, tRNA helps to arrange the amino acids in their proper sequence for the synthesis of a protein. Base pairing between the anticodon on the tRNA (the anticodon is called translation) and the complementary (matching) codon on a mRNA molecule brings the correct amino acid into the growing polypeptide chain.


 mRNA is Made from DNA in a Process called "Transcription"

The manufacture of DNA into RNA takes place only in the nucleus.Transcription is the name of the step in which DNA is copied into mRNA. Transcription is basically the process of rewriting the DAN code in a different nucleic acid language, that of its cousin, mRNA.  It's job is to bring the DNA message (how to make protein) out of the nucleus and into the cytoplasm, where the protein-making "machinery" (ribosomes) are located. 

The process of Transcription

1. Before transcription can begin, the portion of the DNA double helix to be transcribed must be "UNZIPPED" by separating the two strands at their hydrogen bonds. This is done by an enzyme named HELICASE.  Where helicase has separated the two DNA "backbones" there are now rows of nitrogenous bases just waiting to be "read" and "rewritten" into another language: mRNA 

2. Only one of the DNA strands in the double helix is read and used as template to manufacture mRNA. Name of strand of DNA that is transcribed to create m-RNA is antisense strand. The enzyme that attaches to the unzipped DNA and creates a complementary strand of mRNA is called RNA POLYMERASE (see figure below).

3. The Final Product of transcription is a messenger RNA. It resembles half of a DNA molecule and carries the "message" containing instructions for protein synthesis from the DNA "gene" in the nucleus to the ribosomes in the cytoplasm.

The process of transcription.


Synthesis of protein using  genetic message carried by mRNA

Translation is the process of synthesis of a protein by ribosomes, using mRNA as a template.

The process of making protein from mRNA.

Interesting facts about RNA

RNA technologies: RNAi and antisense RNA

Brifely, both RNAi (RNA interference) and antisense RNA induce destruction of mRNA in the cytoplasm and inhibit or block production of protein for a particular function (e.g. inducing fruit ripening, causing cancer etc.). 

1. RNAi: Double stranded RNA is introduced into a cell and gets chopped up by the enzyme dicer to form siRNA. siRNA then binds to the RNA-induced silencing complex (RISC) and is unwound. The anitsense RNA complexed with RISC binds to its corresponding mRNA which is then cleaved by the enzyme slicer and makes it inactive (see figure below).
2. Antisense RNA:
Antisense RNA is a RNA strand which has a mirror image of nucleotide bases of a mRNA strand.
When an artificial gene (DNA) is introduced into a cell, it produces an antisense RNA complementary to the cell's own mRNA and forms RNA duplex  with the mRNA (see figure below). The formation of double stranded RNA inhibits gene expression ( = No production of functional protein because protein synthesis requires single stranded mRNA molecule as a template).
RNAi mechanism                Antisense RNA mechanism
                               
Applications

Cancer gene therapy:
RNAi and antisense RNA technologies have been used in reducing expression of cancer causing genes in  human cancer cell lines.
Control of fruit ripening: Antisense RNA technology has been used to suppress expression of fruit ripening genes to make the fruits stay longer in vine and extend marketing period. e.g. the flav'r sav'r tomato (the first genetically modified food crop introduced in US market in 1995 by a company called Calgene).

References

Biochemistry & molecular biology of plants. Edited by Bob B. Buchanan, Wilhelm Gruissem, Russell L. Jones. Rockville, Md.  American Society of Plant Physiologists, c2000.

Essentials of Genetics. Second edition. Edited by Williams S. Klug and Michael R. Cummings. Prentice Hall, NJ, 1996.

Encyclopedia of RNA. www.sciencedaily.com/encyclopedia/rna

http://www.sciencedaily.com/encyclopedia/central_dogma_of_molecular_biology

http://www.encyclopedia.com/html/section/nucleica_RNAandProteinSynthesis.asp

www.google.ca/image search 

http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/AntisenseRNA.html

 http://www.ambion.com/techlib/resources/RNAi/

http://dragon.zoo.utoronto.ca/~jlm-gmf/T0501D/results_index.html

http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/AntisenseRNA.html

http://en.wikipedia.org/wiki/RNA_world_hypothesis

http://www.newton.dep.anl.gov/askasci/mole00/mole00331.htm

http://dwb.unl.edu/Teacher/NSF/C08/C08Links/www.iacr.bbsrc.ac.uk/notebook/courses/guide/rnast