Tuesday, March 27, 2012

Daily Newsletter March 27, 2012

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Daily Newsletter March 27, 2012

Today's Topic:
Transcription Translation is the genetic process where a single strand of DNA acts as a template for the construction of a complementary RNA strand. Generally when talking about transcription, we will be talking about the formation of messenger RNA (mRNA), which carries the code for one gene to a ribosome where it is translated into a protein.

DNA holds the "permanent" copy of the genes needed to make a functional organism. Think of DNA as a locked safe where you hold all your company's blueprints, patents and documented procedures. You don't want to loose these, or risk that they might be changed. You only bring them out to make copies of them, then they go back to the safe. This is what happens with your DNA. You keep it tightly locked up (in a double-helix that is coiled around histones, and then possibly supercoiled), and open it up only when you NEED to make a copy. Notice how NEED is highlighted? Do you think it might be an important concept?

In eukaryotic DNA every gene starts with a promoter. This is a sight of ~8 nucleotides visible in the major groove of DNA. The transcription complex recognizes this sequence as a "START" indicator. The main core of the transcription complex will be RNA polymerase. This enzyme works to build a strand of RNA complementary to DNA. The name polymerase indicates that it is involved with dehydration synthesis polymerazation reactions (taking one nucleotide, and adding it to a growing chain of nucleotides). Like DNA polymerase, RNA polymerase builds in the 5' to 3', and builds phosphodiester linkages between nucleotides.

But RNA polymerase can not act alone. In eukaryotic systems, initiation factors are needed to recognize the promoter region, and then to correctly align the RNA polymerase. Below is a great picture showing the initiation complex and the RNA polymerase II holozyme (RNA polymerase II with all associated protein structures).
As you can see, TATA Binding Protein (TBP) is the first structure to attach to DNA.  It recognizes the TATA sequence in the major groove of the DNA double helix.  It then forces the the DNA to bend, and acts as a signal to other enzymes directing interactions with DNA.  A cascade of reactions occur to then produce the Preinitiation Complex, which ensures that the transcription complex is positioned correctly over the Transcription Start Site, and begins the unwinding (sometimes referred to as denaturation) of the double helix.  The Transcription Complex then begins to read the template strand of DNA, and makes an RNA copy (Elongation).

NOTE:  Elongation works due to base complementarity.  Ribonucleotide triphophates are brought into the transcription complex, and are added to the free 3' end of the growing RNA strand.

Termination:  We are not going to spend a lot of time on termination.  There are a couple different models of eukaryotic transcription termination.  The main feature is that there is a signal sequence of deoxyribonucleotides in DNA that signals the end of transcription.

mRNA processing:  Once transcription is complete, in eukaryotes, the RNA needs to be processed.  The following is a quick reference for mRNA processing:
  • 5' capping:  To protect the mRNA from ribonucleases (RNA degrading enzymes) that attack the 5' end, 7-methylguanosine is added to the 5' end.  Usually, the 5' ribonucleotide is replace by this compound.  Additionally, methyl groups can be added to the sugar-phosphate backbone to further protect the mRNA.
  • Polyadenylation:  In maturing RNA to mRNA, a poly-A tail is added (usually after cleaving off a small section of the 3' end).  This process adds ~250 adenyls to the 3'end of the molecule.  This is needed to stabilize the molecule and facilitate export through the nuclear pores.  As mRNA is translated, the poly-A tail gets shorter.  When short enough, the mRNA is degraded.  Thus, the polyadenylation (poly-A tail) is responsible for setting a time limit to the mRNA.
  • Splicing:  The RNA is composed of both coding (exon) and non-coding (intron) regions.  To mature into mRNA, the introns have to be removed, and the remaining exon spliced together.  This job is the responsibility of the splicosomes. 
    • The above image is a quick reference to the effects of splicing.

      • The above image is a quick reference to the effects of the splicosome.
Once RNA has been processed (matured), it is ready to be used in translation (protein synthesis).


Daily Challenge: Transcription In your own words, discuss the process of transcription, and the formation (maturation) of mRNA. Remember that we have focused on eukaryotic transcription. Briefly, how does prokaryotic (specifically bacterial) transcription differ from eukaryotic transcription?

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