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Eukaryotic transcription is the elaborate process that eukaryotic cells use to copy genetic
information stored in DNA into units of RNA replica. A eukaryotic cell has a nucleus that
separates the processes of transcription and translation. Eukaryotic transcription occurs within
the nucleus where DNA is packaged into nucleosomes and higher order chromatin structures.
The complexity of the eukaryotic genome necessitates a great variety and complexity of gene
expression control.
Eukaryotes have three nuclear RNA polymerases, each with distinct roles and properties
RNA polymerase I
RNA polymerase I (Pol I) catalyzes the transcription of all rRNA genes except 5S rRNA.
These rRNA genes are organized into a single transcriptional unit and are transcribed into
a continuous transcript.
This precursor is then processed into three rRNAs: 18S, 5.8S, and 28S. The transcription
of rRNA genes takes place in a specialized structure of the nucleus called the nucleolus,
where the transcribed rRNAs are combined with proteins to form ribosomes.
Initiation
Termination
The primary transcript is processed into the mature 18S, 5.8S and 28S rRNAs. The processing
involves exo- and endo-nucleolytic cleavages guided by snoRNA (small nucleolar RNAs) in
complex with proteins. The mature rRNAs contain modified nucleotides which are added after
transcription by a snoRNA-dependent mechanism.
5S ribosomal RNA is transcribed by RNA polymerase III in the nucleoplasm. Each eukaryotic
cell contains a high number of copies of the 5S coding gene (up to 20 000 copies per cell). 5S
rRNA contains overlapping binding sites for two different proteins, ribosomal protein L5 and
transcription factor TFIIIA. The mutual exclusive binding of these two proteins to 5S rRNA is
important for coordinating the expression of 5S rRNA to the production of the other rRNAs.
RNA polymerase II (RNAP II and Pol II) is an enzyme found in eukaryotic cells. It
catalyzes the transcription of DNA
to synthesize precursors of mRNA
and most snRNAs, siRNAs, and
all miRNAs and microRNA. A
550 kDa complex of 12 subunits,
RNAP II is the most studied type
of RNA polymerase. A wide range
of transcription factors are required for it to bind to upstream gene promoters and begin
transcription.
Many Pol II transcripts exist transiently as single strand precursor RNAs (pre-RNAs) that
are further processed to generate mature RNAs. For example, precursor mRNAs (pre-
mRNAs) are extensively processed before exiting into the cytoplasm through the nuclear
pore for protein translation.
Initiation
Processing of mRNA
The steps:
A 5' cap (also termed an RNA cap, an RNA 7-methylguanosine cap, or an RNA m7G
cap) is a modified guanine nucleotide that has been added to the "front" or 5' end of a
eukaryotic messenger RNA shortly after the start of transcription. The 5' cap consists of a
terminal 6-methylguanosine residue that is linked through a 5'-5'-triphosphate bond to the
Splicing is the process by which pre-mRNA is modified to remove certain stretches of non-
coding sequences called introns; the stretches that remain include protein-coding sequences and
are called exons. Sometimes pre-mRNA messages may be spliced in several different ways,
allowing a single gene to encode multiple proteins. This process is called alternative splicing.
Splicing is usually performed by an RNA-protein complex called the spliceosome, but some
RNA molecules are also capable of catalyzing their own splicing.
Editing
Polyadenylation
Polyadenylation occurs during and immediately after transcription of DNA into RNA. After
transcription has been terminated, the mRNA chain is cleaved through the action of an
endonuclease complex associated with RNA polymerase. After the mRNA has been cleaved,
around 250 adenosine residues are added to the free 3' end at the cleavage site. This reaction is
catalyzed by polyadenylate polymerase. Just as in alternative splicing, there can be more than
one polyadenylation variant of an mRNA.
Polyadenylation site mutations also occur. The primary RNA transcript of a gene is cleaved at
the poly-A addition site, and 100-200 A’s are added to the 3’ end of the RNA. If this site is
altered, an abnormally long and unstable mRNA results. Several beta globin mutations alter this
site: one example is AATAAA -> AACAAA. Moderate anemia was result.
RNA polymerase III (Pol III) transcribes small non-coding RNAs, including tRNAs, 5S rRNA,
U6 snRNA, SRP RNA, and other stable short RNAs such as ribonuclease P RNA.
RNA Polymerases I, II, and III contain 14, 12, and 17 subunits, respectively.
All three eukaryotic polymerases have five core subunits that exhibit homology with the
β, β’, αI, αII, and ω subunits of E. coli RNA polymerase.
An identical ω-like subunit (RBP6) is used by all three eukaryotic polymerases, while the
same α-like subunits are used by Pol I and III.
The three eukaryotic polymerases share four other common subunits among themselves.
The remaining subunits are unique to each RNA polymerase. The additional subunits
found in Pol I and Pol III relative to Pol II, are homologous to Pol II transcription factors.
Crystal structures of RNA polymerases I and II provide an opportunity to understand the
interactions among the subunits and the molecular mechanism of eukaryotic transcription
in atomic detail.
Initiation: the construction of the polymerase complex on the promoter. Pol III is unusual
(compared to Pol II) requiring no control sequences upstream of the gene, instead normally
relying on internal control sequences - sequences within the transcribed section of the gene
(although upstream sequences are occasionally seen, e.g. U6 snRNA gene has an upstream
TATA box as seen in Pol II Promoters).
Class I
Class II
Class III
Termination
Polymerase III terminates transcription at small polyTs stretch (5-6). In Eukaryotes, a hairpin
loop is not required, as it is in prokaryotes
1. tRNA is transcribed by RNA polymerase III. The transcription product, the pre-tRNA,
contains additional RNA sequences at both the 5’ and 3’-ends. These additional sequences are
removed from the transcript during processing.
The additional nucleotides at the 5’-end are
removed by an unusual RNA containing enzyme
called ribonuclease P (RNase P).
4. Mature tRNAs can contain up to 10% bases other than the usual adenine (A), guanine (G),
cytidine (C) and uracil (U). These base modifications are introduced into the tRNA at the final
processing step. The biological function of most of the modified bases is uncertain and the
translation process seems normal in mutants lacking the enzymes responsible for modifying the
bases.