UNIVERSITY OF CALOOCAN CITY

Biglang Awa St., Corner Catleya St., EDSA, Caloocan City

COLLEGE OF EDUCATION

GENETICS
SUBJECT CODE: MBS 421
TOPIC OR LESSON: RNA MOLECULES AND RNA PROCESSING
WEEK: 13
SUB-TOPIC/S: 1. Genes complex structure
2. Messenger RNAs, Transfer RNAs, and Ribosomal
RNAs

OVERVIEW OF THE TOPIC

Imagine that you run a book-making factory, and you've just printed up
all the pages of your favorite book. Now that you have the pages, is the book
ready to go? Well...books usually have front and back covers. So, you might
want to put those on. Also, were there any blank or messed-up pages made
during printing? You should probably check for those and remove them
before selling your books, or you might end up with some unhappy
customers.

The steps we just talked about are pretty similar to what happens to
RNA transcripts in the cells of your body. In humans and other eukaryotes, a
freshly made RNA transcript (hot off the RNA polymerase "presses") is not
quite ready to go. Instead, it's called a pre-mRNA and has to go through
some processing steps to become a mature messenger RNA (mRNA) that can
be translated into a protein

Ribonucleic acid (RNA) is a molecule that is present in the majority of

living organisms and viruses. It is made up of nucleotides, which are ribose
sugars attached to nitrogenous bases and phosphate groups. The
nitrogenous bases include adenine, guanine, uracil, and cytosine. RNA
mostly exists in the single-stranded form, but there are special RNA viruses
that are double-stranded. The RNA molecule can have a variety of lengths
and structures. An RNA virus uses RNA instead of DNA as its genetic material
and can cause many human diseases. Transcription is the process of RNA
formation from DNA, and translation is the process of protein synthesis from
RNA. The means of RNA synthesis and the way that it functions differs
between eukaryotes and prokaryotes. Specific RNA molecules also regulate
gene expression and have the potential to serve as therapeutic agents in
human diseases.

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 LEARNING
OUTCOMES

1. Discuss the concepts of collinearity

2. Differentiate the different types of RNA
3. Identify the structure of ribosomes

LEARNING
OBJECTIVES
At the end of the lesson, students should be able to:

a. Being able to identify the different types of RNA

b.
c.

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 EXPLAIN

RNA MOLECULES

RNA PROCESSING

In eukaryotic cells, before RNA polymerase II-generated transcripts could be

translated into protein products, these transcripts (pre-mRNAs) need to be
suitably processed to form messenger RNA (mRNA). Three major events
constitute pre-mRNA processing: (a) 5′-end capping, (b) splicing, and (c) 3′-
end polyadenylations. In 5′-capping, the 5′-triphosphate of the nascent
transcript is hydrolyzed to a diphosphate and a guanosine monophosphate is
added in a reverse 5′-5′ orientation. Subsequently, the GpppN- cap is
methylated to form m7GpppN-. In splicing, the noncoding sequences
(introns) that separate the coding sequences (exons) are removed and the
exons are joined together. 3′-Polyadenylation involves cleavage of the
transcript at a specific site and addition of a poly(A) tail. In addition, there is
a fourth mechanism of RNA processing, known as RNA editing, in which
instead of stretches of the transcript, individual bases are inserted, deleted,
or altered post-transcriptionally.
Three Main Types of RNA are involved in Protein Synthesis:
Pre-messenger RNA (pre-mRNA)
Are the immediate products of transcription in eukaryotic cells. Pre-mRNAs
are modified extensively before becoming mRNA and exiting the nucleus for
translation to protein.
Messenger RNA
Messenger RNA a type of RNA found in cells. mRNA molecules carry the
genetic information needed to make proteins. They carry the information
from the DNA in the nucleus of the cell to the cytoplasm where the proteins
are made.

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Transfer RNA
Transfer RNA is a small RNA molecule that plays a key role in protein
synthesis. Transfer RNA serves as a link (or adaptor) between the messenger
RNA (mRNA) molecule and the growing chain of amino acids that make up a
protein.
Ribosomal RNA
A type of stable RNA that is a major constituent of ribosomes. It ensures the
proper alignment of the mRNA and the ribosomes during protein synthesis
and catalyzes the formation of the peptide bonds between two aligned amino
acids during protein synthesis

Key points:

• When an RNA transcript is first made in a eukaryotic cell, it is

considered a pre-mRNA and must be processed into a messenger RNA
(mRNA).
• A 5’ cap is added to the beginning of the RNA transcript, and a 3’ poly-
A tail is added to the end.
• In splicing, some sections of the RNA transcript (introns) are removed,
and the remaining sections (exons) are stuck back together.
• Some genes can be alternatively spliced, leading to the production of
different mature mRNA molecules from the same initial transcript.

In eukaryotic cells, pre-mRNAs undergo three main processing

steps:
• Capping at the 5’ end
• Addition of a poly(A) tail at the 3’ end
• Splicing to remove introns

Capping
In the capping step of mRNA processing, a methylated-guanosine (7-methyl-
G) is linked to the phosphates at the 5’ end of the mRNA. The cap protects

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the 5’ end of the mRNA from degradation by nucleases and also helps to
position the mRNA correctly on the ribosomes during protein synthesis

Poly(A) tail addition

The 3’ end of a eukaryotic mRNA is first trimmed, then an enzyme called

Poly(A) Polymerase adds a “tail” of about 200 ‘A’ nucleotides to the 3’ end.
There is evidence that the poly(A) tail plays a role in the efficient translation
of the mRNA, as well as in the stability of the mRNA. The cap and the poly(A)
tail on an mRNA indicate that the mRNA is complete (i.e., not defective).

Splicing
In many cases, splicing now appears to occur as the transcript is being
synthesized. Introns are removed from the pre-mRNA by the activity of a
complex called the spliceosome. The spliceosome is made up of proteins and
small nuclear RNAs (snRNAs) that associate to form protein-RNA enzymes
called small nuclear ribonucleoproteins or snRNPs (pronounced SNURPS). The
splicing machinery must be able to recognize sequences that are specific to

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splice junctions (i.e., the end of each exon and the start of the next) in order
to correctly cut out the introns and join the exons together to make the
mature, spliced mRNA.

GENES COMPLEX

MESSENGER RNA

- Messenger RNA (abbreviated mRNA) is a type of single-stranded

RNA involved in protein synthesis. mRNA is made from a DNA template
during the process of transcription.
- The role of mRNA is to carry protein information from the DNA in a
cell’s nucleus to the cell’s cytoplasm (watery interior), where the protein-
making machinery reads the mRNA sequence and translates each three-base
codon into its corresponding amino acid in a growing protein chain.
- Messenger RNA or mRNA. So, mRNA really is a form of nucleic acid,
which helps the human genome which is coded in DNA to be read by the
cellular machinery. So, we have DNA in our nuclei.
- And then we have ribosomes and other cellular organelles which
translate DNA. But between the DNA code itself, and the machinery that uses
DNA to make proteins, there has to be a translator.
- And mRNA is actually the translated form of DNA that the machinery
can recognize and use to assemble amino acids into proteins. So this is really
a fundamental link between what we think of as being the code of life and
the actual cell being able to construct a living organism.
- And in that sense, although DNA gets discussed a lot more than RNA,
mRNA is a really crucial piece of the fundamental way in which the living
organism is created.

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TRANSFER RNA

RIBOSOMAL RNA

Functions of Ribosomal RNA

The primary function of rRNA is in protein synthesis – in binding to

messenger RNA and transfer RNA to ensure that the codon sequence of the
mRNA is translated accurately into an amino acid sequence in proteins. To
achieve this, rRNA has a distinctive three-dimensional shape involving
internal loops and helices that create specific sites within the ribosome – the
A, P, and E sites. The P site is for binding a growing polypeptide, the A site
anchors an incoming tRNA charged with an amino acid. After peptide bond
formation, the tRNA binds briefly to the E site before leaving the ribosome. In
addition, rRNA also has sites for binding to some ribosomal proteins and
careful analysis has demarcated the exact residues in both the RNA and
protein. Ribosomal RNA is also expressed in every cell of all extant species.
The sequence of the core catalytic sites is also highly conserved making
rRNA an excellent tool for the study of taxonomy and phylogenetics. There is
a difference in the rate of evolution of residues on the surface and interior of
rRNA, and nucleotides involved in core catalytic activity, such as in the
formation of a peptide bond, appear to have predated the appearance of life
on earth. The extent to which two species differ in rRNA sequences can give
a good estimate of their evolutionary distance. Many antibiotics target

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prokaryotic rRNA and recently the binding sites for antibiotics such as
streptomycin and tetracycline on rRNA have been indicated. It has also been
shown that antibiotic resistance often stems from point mutations in these
binding sites. For instance, the resistance of Euglena and E. coli to
streptomycin stems from a mutation in the 16S rRNA sequence. Similar
results were found for the resistance of Streptomyces to Spectinomycin.
Tetracycline resistance appears to come from mutations in the 30S rRNA. In
a new dimension to the function of rRNA, its precursors (preribosomal RNA)
have been implicated in the generation of micro-RNA that mediate
inflammation and cardiac disease in response to mechanical stress. The
mechanisms of this activity are still being elucidated.

Role of Ribosomal RNA in Translation

Translation of the mRNA sequence requires the involvement of rRNA at every

step – initiation, elongation, and termination.

Messenger RNAs carry the genetic information coded in the DNA into the
cytoplasm where the nucleotide sequence is read by ribosomes in stretches
of three bases called codons. Four nucleotides, Adenine, Uracil, Guanine and
Cytosine, can be arranged to form a total of sixty-four triplet codons. Each
codon corresponds to a single amino acid and thus codes for the protein
sequence.

Prokaryotic translation begins with the 16S rRNA base pairing with the Shine-
Dalgarno consensus sequence in mRNA. Since the Shine-Dalgarno sequence
is 6-10 nucleotides upstream of the start codon, binding with rRNA allows the
start codon to be positioned within the ribosome. This interaction is mediated
by other proteins, which also recruit the larger ribosomal subunit and
subsequently, the first codon is translated. In eukaryotes, eukaryotic
Initiation Factors 4E and 4G (eIF4E and eIF4G) bind to the 5′ ends of the
mRNA, recruiting both the smaller subunit of a ribosome and a tRNA carrying
methionine. The ribosome scans the mRNA to locate the start codon, after
which the initiation factors dissociate from the translation machinery.

Every new amino acid, attached to a tRNA, arrives at the A site. Base pairing
between the codon on the mRNA and the complementary anticodon on the
tRNA changes the conformation of three residues on the 16S rRNA. These
residues interact with the anticodon, stabilize the tRNA-mRNA complex and
enzymatic activity of the rRNA positions the aminoacyl-tRNA fully within the
A site.

The polypeptide that has been synthesized so far is bound to the P site on
the ribosome. Ribosomal RNA in the larger subunit catalyzes the reaction

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that forms a peptide bond between the amino acid in the A site and the
growing polypeptide chain in the P site. Polypeptide synthesis is terminated
when the ribosome reaches a stop codon, and rRNA catalyzes the addition of
a water molecule to the polypeptide in the P site.

ELABORA
TE
Students will be asked to make a table which shows the difference of the three types of RNA

EVALUAT
E

REFERENC
ES

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 ADDITIONAL
MATERIALS

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