01.

DNA Structure and Organization

Griffith 1928 Transformation experiments

Hershey and Chase 1952 - Transduction

DNA is a polar polymer 5 Ph4 3 OH

Informao adicional: 1. Densidade da molcula nativa muito alta 2. Decomposio da molcula nativa em nucletidos resultava em que as propores dos diferentes componentes eram sempre: Guaninas = Citocinas Adeninas = Timinas 3. A molcula nativa tinha um dimetro constante (20 )

Estabilidade do DNA

Interaes hidrofbicas entre as bases que resultam na sua orientao principalmente perpendicular ao eixo da molcula.

Desnaturation e renaturation do DNA

DNA melting curves

Southern blotting

Southern blotting

Estrutura secundaria do DNA

Regio de dupla simetria com sequencias repetidas invertidas

Linear DNA molecule

Link number = 0 Positive supercoiling Link number = 3

Nucleossoma

Modelos da organizao da cromatina Bead on a string 11 nm

Modelos da organizao da cromatina Solenoid 30 nm

Remodelao da cromoatina

Dependente do ATP

Diversas protenas interatuam com o DNA

C-terminal das histonas

O Cdigo epigentico

O Cdigo epigentico

Organizao do DNA nu ncleo

Packing ratio

6-7

40

680

1.2 x 10

The chromosome Scaffold

Remover DNA com nucleases e DNA com NaCl Protenas no histonas Topoisomerase II Structural Maintenance of Chromosomes SMCs

Scaffold Mitotic

Matrix Interphase (nucleus)

Codensin is involved in the organization of mitotic chromosomes

Cohesin maintains sister chromatids together from G1 to end of Mitosis

DNA Condensin Cohesin

Cornelia de Lange Syndrome (1/10,000) severe developmental deffects Mutations at SMC1/3 or SCC2 (chaperone of SCC1)

Genoma de um organismo Chromosome painting

Control 1 N = Haploide 2 N = Diploide

cancer cell

Chromosome territories in human cells

Gene localization changes within the nucleous in response to transcription

Genomics

DNA complexity

25,000

Tipos de sequencias no genoma

TE

Repeats

Transpose

Repeat

Transposons

Barbara McClintock 1902-1992 Nobel prize of Physiology and Medicine 1983

Transposition

-Normalmente reprimida -Pode ser activada -Induz alteraes em genes -Induz quebras cromossomicas

Transposons
copy paste via RNA or DNA

Transposes podem inativar genes

Heterocromatina

O Genoma humano

LINEs: Long Interspersed Nuclear Elements - Heterogenous in lenght - Transcribed by RNA pol II - Code for reverse transcriptase and endonuclease - 500,000 copies - Move by coping so enlarge the genome

SINEs: Short Interspersed Nuclear Elements - Short <500 b - Transcribed by RNA pol III - Non-coding - Alu sequences (280 bp) cut RE Alu - 1,5 M copies - Use TEs for mobilization