Basic Molecular Genetics

Dr. Upik A. Miskad, PhD Unhas Research Center

Medical genetics in the past : Minor subject in the school. Related to uncommon diseases. (ex/ hemofilia, sindr.down, buta warna) Only specialist and affected patients concern to the subject. Medical genetics today : Explain underlying molecular pathobiology of many diseases. (mjelaskan semua pyk kec lingk) Mainstream of medical science and public health. Powerful tool to establish: diagnosis, classification, prognosis, prediction of complication, as well as response to the treatment.(alat utk mnegakkan diag, etc)

Dr. Upik A. Miskad, PhD Unhas Research Center

CELLS

Cells are the basic units of living organisms. Although a nerve cell looks entirely different from a red blood cell, their organizations are essentially the same. Cells (Prokariotik dan eukaryotik)

Eukaryote versus prokaryote

Circular shape of microbial DNA

Cells

* building block of all life

* very small : - membrane - cytoplasm : RNA - nucleus

chromosomes DNA

 An organism may contain

many types of somatic cells, each with distinct shape and function.
However, they all have the

same genome.

 The genes in a genome do not

have any effect on cellular functions until they are "expressed".

Different types of cells express

different sets of genes, thereby exhibiting various shapes and functions.

The information goes only one way

The central dogma states that once information has passed into protein it cannot get out again. The transfer of information . from nucleic acid to nucleic acid, or from nucleic acid to protein, may be possible, but transfer from protein to protein, or from protein to nucleic acid, is impossible.

GENE

By definition, a gene includes the entire nucleic acid sequence necessary for the expression of its product (peptide or RNA). Such sequence may be divided into regulatory region and transcriptional region.

ASAM NUKLEAT
Asam Deoksiribonukleat DNA (Deoxyribonucleid Acid) Asam Ribonukleat RNA (Ribonucleid Acid)

DNA
DNA adalah suatu

polimer yang panjang tidak bercabang, mengandung 2 rantai polynucleotida, yang tersusun sebagai suatu bentuk antiparalel yang mengelilingi aksis.

DNA
DNA tersusun sebagai

suatu right-handed double helix dengan kelompokan gulaposfat sebagai tulang punggungnya dan basa basa tersusun dibagian pinggirnya

Nucleotides (basa):

A - T
G - C

Sugar: De oxy ribose (DNA) Ribose (RNA)

Phosphate:

DNA
* can be renatured and denatured * denatured : boils temperature extreme of PH (PH < 3 PH > 10) * renatured : 650 C

BASES
There are five different bases, each is denoted by a

single letter as given in the parenthesis: Adenine (A), Cytosine (C), Guanine (G), Thymine (T), and Uracil (U). Among them, A, C, G and T exist in DNA; A, C, G and U exist in RNA.

Setiap nucleotida disusun oleh 3 komponen: 1. Gula/ pentose, 2. Basa/ base 3. Phosphate group. Nucleosida Disusun oleh gula/ pentosa dan basa saja

Basa berikatan dengan dengan Deoxyribosa dengan Glycocyl linkage (warna ungu) membentuk NUCLEOSIDA. Gugus gula pada ikatan basa dan gula bila mengalami phosphorilasi (penambahan gugus phospat) akan mementuk NUCLEOTIDA.

Phosphodiester bridge

Nucleotida dihubungkan

satu sama lain oleh jembatan phosphodiester (Phosphodiester bridge)

Yang menghubungkan

antara gula pada nucleotida yang satu dengan phosphat pada nucleotida yang lainnya

STRUKTUR KIMIA GULA/ PENTOSA

The chemical structure of pentose which contains five carbon atoms, labeled as C1' to C5'. The pentose is called ribose in RNA and deoxyribose in DNA, because the DNA's pentose lacks an oxygen atom at C2'.

PENTOSA/GULA
RNA : RiboNucleid Acid

DNA: DeoxyriboNucleid Acid

BASA
Ada 2 macam Basa pada Asam nukleat: Purin Pirimidin Purin: Struktur kimianya terdiri dari 2 bergandengan Terdiri dari 2 macam:
A: Adenine G: Guanine

cincin yang

Pirimidin: Struktur kimianya terdiri hanya 1 cincin. Terdiri dari 3 macam:

C: Cystosine T: Thymine U: Urasil

STRUKTUR KIMIA BASA

Ada 2 macam Basa pada Asam nukleat: 1. Purin 2. Pyrimidin Purin: A: Adenine G: Guanine

Pirimidin: C: Cystosine T: Thymine U: Urasil

BASA YANG MEMBENTUK DNA

Computer model of base pairing in DNA.
In a normal DNA molecule,

Adenine (A) is paired with Thymine (T), Guanine (G) is paired with Cytosine (C).
The uracil (U) of RNA can also

pair with adenine (A), since U differs from T by only a methyl group located on the other side of hydrogen bonding (ikatan hidrogen)

FUNGSI BASA DALAM DNA

Utk : Mempertahankan kestabilan DNA

dengan adanya ikatan hidrogen antara pasangan pasangan basa dari 2 rantai polinucleotida dalam arah horisontal.
A DNA molecule has two strands, held

together by the hydrogen bonding between their bases. As shown in the figure: Adenine can form two hydrogen bonds with Thymine; Cytosine can form three hydrogen bonds with Guanine.

DNA Double Helix

The two strands form a "double

helix" structure, which was first discovered by James D. Watson and Francis Crick in 1953. In this structure, also known as the B form, the helix makes a turn every 3.4 nm, and the distance between two neighboring base pairs is 0.34 nm. Hence, there are about 10 pairs per turn. The intertwined strands make two grooves of different widths, referred to as the major groove and the minor groove, which may facilitate binding with specific proteins.
The normal right-handed "double helix" structure of DNA, also known as the B form.

RNA (RIBONUCLEID ACID)

Ribonucleid Acid mempunyai struktur kimia yang mirip DNA, kecuali:

1. Gulanya ribosa 2. Basa thymine (T) diganti dengan Uracyl (U) 3. merupakan single strand (rantai tunggal

RNA (RIBONUCLEID ACID)

Most cellular RNA molecules are single stranded. They may form secondary structures such as stem-loop and hairpin.

Ada 3 macam molekul RNA

1. Messenger RNA (mRNA) RNA yang mengarahkan pembentukan molekul protein. 2. Transfer RNA (tRNA) RNA yang berfungsi membawa Asam Amino ketempat pembentukan protein. 3. Ribosomal RNA (rRNA) RNA yang berfungsi membentuk komponen ribosom.

RNA (RIBONUCLEID ACID)

Molekul RNA disintesa dari DNA melalui proses yang disebut DNA Transcription.

Molekul RNA relative lebih pendek dibandingkan dengan DNA karena RNA dicopy dari satu daerah yang terbatas.

mRNA
Molekul RNA disintesa

dari DNA melalui proses yang disebut DNA Transcription. mRNA membawa informasi untuk sintesa protein Perhatikan tidak ada T pada RNA, T (thymin) diganti dengan U (Urasil)

CENTRAL DOGMA
Essential steps involved in the expression of protein genes

The central dogma According to the process, the flow of genetic information is in the following direction:

DNA > RNA > Protein.

GENE EXPRESSION
Gene expression" means the production of a protein or a functional RNA from its gene. Several steps are required:

Transcription: A DNA strand is used as the template to

synthesize a RNA strand, which is called the primary transcript. RNA processing: This step involves modifications of the primary transcript to generate a mature mRNA (for protein genes) or a functional tRNA or rRNA. For RNA genes (tRNA and rRNA), the expression is complete after a functional tRNA or rRNA is generated. However, protein genes require additional steps: Nuclear transport: mRNA has to be transported from the nucleus to the cytoplasm for protein synthesis. Protein synthesis: In the cytoplasm, mRNA binds to ribosomes, which can synthesize a polypeptide based on the sequence of mRNA.

PROSES TRANSKRIPSI DNA

1. Pengkopian seluruh panjang DNA yang diinginkan untuk dikopi Meliputi ekson dan intron dengan bantuan enzym RNA polymerase membentuk primary transkript

PROSES TRANSKRIPSI DNA 2. Sebelum molekul RNA meninggalkan nukleus,

enzym pembentuk Komplex RNA (Complex of RNA processing enzyme) akan menghilangkanSemua intron

PROSES TRANSKRIPSI DNA

3. Setelah proses Splicing, RNA yang lebih pendek akan Bergerak ke sitoplasma, untuk selanjutnya membentuk Asam asam amino.

Exon : regio pada RNA yang mempunyai kemampuan mengkode protein Intron : regio pada RNA yang tidak mempunyai kemampuan menkode Protein

 Tiga nucleotida berturut turut dalam mRNA menkode satu asam amino atau stop signal untuk sintesa protein. Tiga rangkaian nucleotida ini disebut CODON

CODON
Adalah suatu sequences yang terdiri dari 3 nukleotida yang mengkode satu asam amino spesifik Jadi seharusnya ada 43=64 asam amino dari 4 kombinasi nukleotida yang ada. Tapi yang hanya ditemukan dalam ptotein adalah 20 asam amino Jadi ada beberapa Codon membentuk asam amino yang serupa.

CENTRAL DOGMA
Essential steps involved in the expression of protein genes

The central dogma According to the process, the flow of genetic information is in the following direction:

DNA > RNA > Protein.

tRNA
Translasi dari mRNA ke protein tergantung pada adaptor molekul yang disebut tRNA. tRNA Berfungsi mengenali dan membawa asam amino yang akan dibentuk. Terdiri atas 80 nukleotida Berbentuk seperti clover leaf

tRNA
Berbentuk tonjolan 3 dimensi. Ada 2 daerah yang penting:

1. anticodon: daerah yang disusun oleh 3 nukleotida yang akan berikatan dengan kodon komplemennya pada mRNA 2. amino acid attachment region: daerah pada tRNA yang akan mengikat asam amino yang dibawanya

PROSES TRANSLASI PROTEIN

Genetic code: A process of translating of the nucleotide sequence into amino acid sequence of protein.
During protein synthesis, the ribosome binds to mRNA and tRNA as shown in the following figure. Only the tRNA containing the anticodon which matches mRNA's codon may join the complex

The mRNA-ribosome-tRNA complex

Pemesanan molekul tRNA pada mRNA memerlukan tempat yang disebut RIBOSOME RIBOSOME: adalah suatu mesin pembentukan protein yang mengandung tRNA yang akan membaca informasi genetik pada mRNA

 PROSES TRANSLASI PROTEIN

pada mRNA-ribosome-tRNA complex

Ribosome akan bergerak sepanjang mRNA dan tRNA yang ada didalamnya akan membaca pesan genetik sambil membawa asam amino yang akan dirangkai dengan asam amino yang lain membentuk polipeptida.

Polipeptida

PROTEIN
Protein dibentuk oleh rangkain polipeptida dan

polipeptida disusun oleh asam amino. Asam asam amino dihubungkan satu sama lain dengan ikatan peptida.

 See you in the next lecture

Thank you