Regulating Transcription - Biochemistry

Transcription is the process of utilizing information in DNA molecules to make RNA molecules. It can occur in eukaryotes (such as humans) and in prokaryotes (such as bacteria). In eukaryotic transcription, the DNA is transcribed to RNA inside the nucleus. Upon completion, the synthesized RNA undergoes further post-transcriptional modifications such as addition of methyl cap, poly-A tail, and removal of introns. After these modifications, the RNA molecule leaves the nucleus, enters the cytoplasm, and undergoes translation (process of synthesizing proteins).

In contrast, bacterial transcription occurs in the cytoplasm and does not involve any of the post-transcriptional modifications. As a result, the transcribed RNA can immediately be used to synthesize proteins; therefore, translation immediately follows transcription in bacteria.

Recall that reverse transcriptase is a special enzyme that converts RNA to DNA (‘reverse’ of transcription). It is found in some viruses such as HIV.

Transcription is the process of transcribing RNA molecules from DNA. This is a normal cellular process that is required for cells to grow and function properly (because these RNA molecules are eventually converted to proteins, the building blocks of cells). Growth of cells occurs in G1 and G2 phases; therefore, transcription occurs during both of these phases.

Note that DNA replication occurs during S phase; therefore, no DNA molecules will be available for transcription during S phase and transcription will be halted.

Sigma factor is a special molecule in bacteria that is used to initiate transcription. In a bacterial cell, RNA polymerase is typically kept in its inactive form, called holoenzyme. When it is needed for transcription, RNA polymerase is converted to its active form by sigma factor. Sigma factor facilitates the binding of RNA polymerase to the gene sequence on the corresponding DNA molecule. Upon binding, RNA polymerase will carry out transcription and generate a new mRNA strand.

Untranslated regions never yield proteins, and thus do not attach to protein kinases. The 5’ end of pre-RNA is capped, and the 3’ end modified by poly A tails. Pre-RNA is, indeed, spliced when introns are removed. This is performed by spliceosomes, which only splice RNA, not DNA.

A Pribnow box is a type of promoter region in bacteria that contains a sequence similar to the eukaryotic TATA box. 5'- TATAAT -3' will be found in the Pribnow box and signifies that the particular section of DNA is a promoter region. The eukaryotic TATA box typically contains the sequence 5'- TATAAA -3' and also serves as a promoter region, typically found upstream of a gene.

Transposable elements are those that can move around within the genome, which increases genetic diversity. Also, due to alternative splicing of introns, multiple distinct proteins can be synthesized from the same exact mRNA transcript. One example of this is antibody production. Segments of DNA can spontaneously switch to become new DNA coding regions (mutation). This also increases genetic diversity, if this occurs in the germ-line cells.

RNA polymerase II is the polymerase that catalyzes the synthesis of mRNA from a coding strand of DNA. Therefore, mRNA synthesis would be greatly affected by an inhibition of RNA polymerase II.

RNA polymerase III catalyzes the synthesis of tRNA - RNA that is responsible for carrying amino acids during translation. So, synthesis of protein will be affected down the line, however the direct effect of an inhibition of RNA polymerase III would be the inability to create tRNA.

For this question, we need to consider how histone acetyltransferases affect histones. Then, we need to determine how these modified histones affects the expression of genes.

First, it's important to note that histones are proteins that mostly contain positive charges. As a result of this, histones are able to associate with DNA very well, since DNA contains a negatively charged backbone. When histones associate with DNA in this way, the DNA molecule becomes tightly coiled around the histones. In this tightly bound conformation, the collection of DNA and proteins are referred to as hererochromatin. What's more is that when the DNA is tightly bound like this, the transcription machinery in the cell is physically blocked from associating with genes. Thus, gene expression is lowered.

Histone acetyltransferases are enzymes that attach acetyl groups to the positively charged lysine residues that are part of histones. Remember, the positive charge of these lysine residues is what allows the histones to associate with the DNA. When acetyl groups are added, the positive charge on these histones becomes neutralized. As a result, the histones are no longer able to associate with the DNA. What this means is that the transcription machinery in the cell is now able to physically access the genes, allowing gene expression to increase.

RNA polymerase travels down DNA beginning at the promoter site (could be TATA box or Hogness box in eukaryotes). It reads the DNA and synthesizes mRNA along the way, until it reaches a point where it reads the DNA and synthesizes a termination sequence. This notifies the RNA polymerase that it should end transcription of the gene.

Spliceosomes recognize the conserved sequence, GU, and splice just before those two nucleotides. They then continue onwards and when they recognize a pyrimidine followed by the nucleotides, AG, they splice again immediately after the AG. This is almost always conserved in introns to ensure proper splicing.

There are few differences between prokaryotes and eukaryotes in what concerns transcription. In prokaryotes there is only one RNA polymerase, while in eukaryotes there are three: I , II and III. In prokaryotes, both sigma factor and rho factor are required for transcription to occur, but not in eukaryotes.