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Harvy Mauricio Velasco P Gentica Humana UNAL

1* 1015 clulas adultas 2*1017 divisiones celulares 6 *109 nucletidos para cada divisin l l celular 1*10 -6 -8 Eficiencia de la l DNA polimerasa li

Proliferacin, Divisin y apoptosis

Noxas externas

Oncogenes y Genes supresores

Reparacin del DNA

UN GEN SE ENCUENTRA AFECTADO

GENERADAS POR LA EVOLUCION


NOVEDAD ADAPTACION ENFERMEDAD MUERTE

ERRORES EN LA REPLICACION
DNA POLIMERASA

TAMAO DE DNA
3.000 3 000 MILLONES DE pb

AGRESION DIRECTA A DNA DEGRADACION INTERNA DE NUCLEOTIDOS

MUTACION Cualquier cambio en el material gentico(DNA nuclear y mitocondrial), que se herede a las generaciones siguientes.

POLIMORFISMO Variacin V i i gentica ti que tiene ms de una variante i t allica en una i secuencia DNA Frecuencia >1%

MUTACIONES GERMINALES
El cambio en el DNA se da en l lnea parental en t l celulas germinales y esta mutacin se HEREDA

MUTACIONES SOMATICAS
Mutacin M t i que no compromete lneas germinales En un tejido especifico

GERMINALES

SOMATICAS

ESPONTANEAS

INDUCIDAS POR EL AMBIENTE O EN EL LABORATORIO

A GRAN ESCALA O MUTACIONES CITOGENETICAMENTE VISIBLES

A PEQUEA ESCALA O CITOGENETICAMENTE INVISIBLES

EXISTE ALTERACION DE ALGUNOS DE LOS COMPONENTES DEL MATERIAL GENETICO O EN EL FLUJO DE INFORMACION DEL DNA HACIA PROTEINAS ERROR EN LA EXPRESION DE LA INFORMACION GENETICA

TIPO SUSTITUCION DELECION INSERCION

DEFINICION Reemplazo de una(s) base(s) por otra(s) Perdida P did o eliminacin d uno o li i i de ms nucletidos Uno o ms nucletidos estn adicionados en una secuencia
Transposicin con duplicacin de material gentico Transposicin sin duplicacin (translocacin)

MUTACIONES

Sustitucin de bases

Inserciones

Deleciones

Transiciones Transversiones

1 o pocos nucletidos

1 o pocos nucletidos

Sustitucin sinnima (neutra) i i ( t ) y no sinnima Conversin gnica, mltiple sustitucin de bases

Expansin de i l tripletas

Grandes deleciones

Otras grandes inserciones: elementos transponibles ibl

En el marco de lectura

Transiciones: Pirimidina por p pirimidina (C-T)


Ms comunes (ISLAS ( CpG)

Transversiones: p p Purina por pirimidina o viceversa

Purina por purina (AG)

SINNIMAS (SILENCIOSAS): Neutral No cambia secuencia a.a. En DNA no codificante Ms comn en DNA codificante En DNA codificante cambia el A.A. en posicin 3 de codn

NO SINNIMAS Altera secuencia TIPOS


Deletreas Sin efecto Efecto benfico

Mayor frecuencia Transversiones que transiciones Mayor frecuencia en secuencias no codificantes Cualquier secuencia puede p tener sustituciones

No sinnimas Sinonimias Sinnimas

Algunas sustituciones en secuencias no codificantes afectan la expresin

p e q n r u s a a e z c r m a M a c M d U a c U o T n a N T S A m O d A q U d C n e C u S e O n C T O d N O m N o S a e N T E S E c U S c V a r E f S a C o A R u m M d T V n N e O o A c O n N S n V T e N E S m A S e S E d p V n E N e A S s S d a o E E p z m f e a r e d x a r o p a e d r n a e 3 a s e a y n p 2 o d g s a n c p o c s a n c c o d n o n c o d o n

SUSTITUCIONES

Gene Histone H3 Histone H4 Actin Aldolase A HPRT Insulin -Globin -Globin Albumin Ig VH Growth hormone Ig Interferon-1 Interferon-

No. of codons 135 101 376 363 217 51 141 144 590 100 189 106 159 136

Nonsynonymous 0.00 0.00 0 00 0.01 0.07 0.13 0.13 0.55 0.80 0.91 1.07 1.23 1.87 2.21 2.79

Synonymous 6.38 6.12 6 12 3.68 3.59 2.13 4.02 5.14 3.05 6.63 5.66 4.95 5.66 5.88 8.59

MUTACIONES QUE GENERAN GANANCIA O PERDIDA DE MATERIAL GENETICO DNA CODIFICANTE Y NO CODIFICANTE INVOLUCRA GENERALMENTE REPETICIONES CORTAS EN TANDEM FENOMENOS DE RECOMBINACION

Short Tandem Repeats (STRs)


AATG

7 repeats 8 repeats

Homologous equal crossover can result in fusion genes. The example shows how intragenic equal crossover occurring between alleles on nonsister chromatids can generate novel fusion genes composed of adjacent segments f t from the t th two alleles. Note that similar exchanges between genes on sister chromatids do not l i i l result in genetic novelty because the gene sequences on the interacting sister chromatids would be expected to be identical.

Unequal crossover and unequal sister chromatid exchange cause insertions and deletions. The examples illustrate unequal pairing of chromatids within a tandemly repeated array. U d Unequal crossover l involves unequal pairing of nonsister chromatids followed by chromatid breakage and rejoining. Unequal sister chromatid exchange involves unequal pairing of sister chromatids followed by chromatid breakage and rejoining. For the sake of simplicity, the breakages of the chromatids are shown to occur between repeats, but of course breaks can occur within repeats. Note that both types of exchange are reciprocal - one of the participating chromatids l ti i ti h tid loses some DNA, while the other gains some.

Unequal crossover in a tandem repeat array can result in sequence homogenization. Note that the initial spread of the novel sequence variant to the same position in the chromosomes of other members of a sexual population p p can result by random genetic drift. Once the mutation has achieved a reasonable population frequency (left panel) it can spread to other positions within the array (right panel). This can occur by successive gain of mutant repeats as a result of unequal crossover (or unequal sister chromatid exchanges) and occasional loss of normal repeats. Eventually the mutant repeat can replace the original repeat sequence at all positions within the array, leading to sequence homogenization for the mutant repeat. Such sequence homogenization is thought to result in species-specific concerted evolution for repetitive DNA sequences. UEC unequal crossover. UEC, l

Tandem gene duplication can result from unequal crossover or unequal sister chromatid exchange, facilitated by short interspersed repeats. The double arrow indicates the extent of the tandem gene duplication f d li ti of a segment t containing gene A and flanking sequences. Original mispairing of chromatids ld b f ili db hi h could be facilitated by a high degree of sequence homology between nonallelic short repeats ( 1, R2). Note that p (R ) the same mechanism can result in large-scale deletions.

Gene conversion involves a nonreciprocal sequence exchange between allelic or nonallelic genes. (A) Interallelic gene conversion. Note the nonreciprocal nature of the sequence exchange - the donor sequence is not altered but the acceptor q p sequence is altered by incorporating sequence copied from the donor sequence. (B) Interlocus gene conversion. This is facilitated by a high degree of sequence homology between nonallelic sequences, as in the case of tandem repeats (C) Mismatch repeats. repair of a heteroduplex. This is one of several possible models to explain gene conversion. The model envisages invasion by one strand of the donor sequence (-) to form a heteroduplex with the complementary (+) ( ) strand of the acceptor sequence, thereby displacing the other strand of the acceptor. Mismatch repair enzymes recognize the mispaired bases in the heteroduplex and correct' the mismatches so that the (+) acceptor sequence i is converted' to b d' be perfectly complementary in sequence to the (-) donor strand. Subsequent replication of the (-) acceptor strand and sealing of nicks results in completion of the conversion.

LUGARES DE DNA
SECUENCIAS CODIFICANTES DEL GEN

CARACTERISTICA

SECUENCIAS INTRAGENIAS NO CODIFICANTES

SECUENCIAS REGULADORAS FUERA DE EXONES

La mayora patolgicas (sustituciones) (1ra y 2da posicn) Sitios mas susceptibles : islas CpG cerca a hotspots Menor porcentaje mutacional (10 15%) (10-15%) Afecta regiones intronicas En las regiones promotoras de los g genes ( (antes del Exon 1) ) Alteracin en la metilacin

Mutations at conserved splice donor (SD) or splice acceptor (SA) sequences (see Figure 1.15 for consensus sequences) result in (A) intron retention where there is failure of splicing and an intervening intron sequence is not g q excised; or in exon skipping where the spliceosome brings together the splice donor and splice acceptor sites of nonneighbouring exons. (B) Sequences that are very similar to the consensus splice donor or splice acceptor sequences may coincidentally exist in introns and exons (sd and sa). These sequences are not normally used in splicing and so are known as cryptic splice sites. A mutation can activate a cryptic splice site by making the sequence more like the consensus splice donor or acceptor sequence and the cryptic splice site can now be recognized and used by the spliceosome (activation of the cryptic splice site). S Fi li i ) See Figures 9.12 and 9 13 9 12 d 9.13 for examples of activation of an exonic and an intronic cryptic splice site, respectively

GANANCIA DE INTRON

PERDIDA DE EXON

GENERACION DE INSERCIONES O DELECIONES EN EL DNA PRODUCIENDO MUTACIONES TIPO FRAMESHIFT

Location and nature of mutation Effect on gene function Extragenic mutation Normally none

Comments Rare mutations may result in inactivation of distant regulatory elements required for normal gene expression (see Figure 8.23)

Multigene deletion Whole gene deletion Whole gene duplication

Abolition

Associated with contiguous gene syndromes (see Figure 16.9)

Abolition Can have effect due to altered gene Large duplications including the peripheral myelin protein 22 gene can cause Charcot-Mariedosage Tooth syndrome (see Figure 16.7)

Whole exon deletion Within exon

Abolition or modification Abolition

May cause shift in reading frame; protein often unstable If loss/change of key amino acids, shift of the reading frame or introduction of premature stop codon If nonconservative substitutions, small in-frame insertions or other mutations at some locations If conservative/silent substitutions or mutation at nonessential sites

Modification

None Whole intron deletion Splice site mutation

None Abolition or modulation of expression Conserved GT and AG signals are critically important for normal gene expression. Mutations ki i t ti may i d induce exon skipping or i t intron retention

Promoter mutation

Abolition or modulation of expression Deletion, insertion or substitution of nucleotides within promoter may alter expression. Complete deletion abolishes function

Mutation of termination codon

Modification

Additional amino acids are included at the end of the protein until another stop codon is reached

Mutation of poly(A) signal

Abolition or modulation of expression Deletion, insertion or substitution of nucleotides within poly(A) site may alter expression. Complete deletion abolishes function

Elsewhere in introns/UTS

Usually none

Change Delete: (i)the (i) the entire gene (ii)partofthegene Insertasequenceintothegene Disrupt the genestructure: (i)by atranslocation (ii)byaninversion (ii) by an inversion Preventthepromoterworking: (i)bymutation (ii)bymethylation Destabilize the mRNA: (i)byapolyadenylationsitemutation (ii)bynonsensemediatedRNAdecay Prevent correct splicing : (i)byinactivatingdonorsplicesite (ii)byinactivatingacceptorsplicesite (iii)byactivatingacrypticsplicesite Introduceaframeshift intranslation Convertacodon intoastopcodon Replace an essential aminoacid Preventpost transcriptionalprocessing Prevent posttranscriptional processing Preventcorrectcellularlocalizationofproduct

Example Mostthalassemia mutations Most thalassemia mutations 60%ofDuchenne musculardystrophy InsertionofLINE1repetitivesequence intoF8C genein hemophiliaA Xautosome translocationsinwomenwithDuchenne musculardystrophy InversioninF8C gene Inversion in F8C gene Globin 29AGmutation FragileXfullmutation(FMR1) globinAATAAAAATAGAmutation Fibrillinmutations(FBN1) PAX3 451+1GTmutation PAX3 4522AGmutation Globin intron 1110GAmutation PAX3 874875insGmutation PAX3 Q254Xmutation PAX3 R271Cmutation Cleavage resistantcollagenN terminalpropeptide in Cleavageresistant collagen Nterminal propeptide in EhlersDanlos VIIsyndrome F508delmutationincysticfibrosis

NOMENCLATURA

EFECTO EN EL ALELO

ALELO NULO

El alelo no genera prod cto producto proteico A. HIPOMORFO Genera una reducida cantidad del producto A.HIPERMORFO Genera una excesiva cantidad del producto p A. NEOMORFO Genera un nuevo producto A. A ANTIMORFO Genera un producto que G d t antagoniza la actividad o funcin d un producto f i de d t normal

Sustitucin a.a. R117H: arginina por histidina hi tidi en 117 (Arg117 (A 117 His) G542X: glicina por Stop (Gly Stop) Sustitucin nucletidos
1162(G>A) 621(G>T) ( )

Deleciones inserciones i i Delta - F508 o F508 nt6232(del5) o nt6232(del ACCTG) ( ) nt409(insC)

IVS4+1G>T :
Cambio de G por T en la primera base del intron 4

409-410insC :
Insercin d C entre l nucleotidos 409 y 410 I i de los l id

PERDIDA DE FUNCION

Enfermedades A.R.

Haploinsuficiencia A.D.

Efecto dominante negativo A.D.

Toxicidad celular

< 50% de la actividad enzimatica

Expansin de tripletas (GRANDES)

GANANCIA DE FUNCION

Protenas hiperfuncionantes o autoestimuladas

Protenas neoformadas

Enfermedades A.D.

Repeticin de tripletas con presencia de protenas con poli glutamina (MEDIANA)

ABL -BCL

Loss of function mutations in the PAX3 gene. The 10 exons of the gene are shown as boxes, with the connecting introns not to scale. The shaded areas show the sequences encoding the two DNA-binding domains of the PAX3 protein. Note that mutations that completely destroy the structure of the PAX3 protein (drawn above the gene diagram) are scattered over at least the first six exons of the gene, but missense mutations (shown below the gene diagram) are concentrated in two regions, the 5 part of the paired domain and the third helix of the homeodomain. A196T is believed to affect splicing. The 874875insG mutation introduces a seventh G into a run of six Gs; it has arisen independently several times and illustrates the relatively high frequency of slipped-strand mispairing

HPRT activity (% of normal) Phenotype >60 860 1.68 1.41.6 14 16 <1.4 Normal Neurologically normal; hyperuricemia (gout) Neurological problem (choreoathetosis) Lesch-Nyhan syndrome ( h h h d (choreoathetosis, h i mutilation) but intelligence normal selflf

Classical Lesch-Nyhan syndrome (MIM 308000; y y ( ; choreoathetosis, self-mutilation and mental retardation)

DELECIONES E INSERCIONES CON FORMACON DE FRAMESHIFT SUSTITUCIONES TIPO NONSENSE MUTACIONES CON ALTERACION EN EL SPLICING

GLOBINA ( talase a) talasemia) DELECION C ON FACTOR 8 (Hemofilia A) INSERCION DISTOFINA (Distrofia de Duchenne) FRAMESHIFT PAX 3 (S W d b ) (S. Waardenburg) I,D,S,Splic

Condition

MIM no.

Gene

Alagille syndrome Multiple exostoses Tomaculous neuropathy Supravalvular aortic stenosis Tricho-rhinophalangeal syndrome Waardenburg syndrome Type 1

118450

JAG1

133700

EXT1

162500

PMP22

185500

ELN

190350

TRPS1

193500

PAX3

Malfunction p Overexpression

Gene PMP22

Disease Charcot-Marie-Tooth disease

MIM no. 118200

Receptor permanently on on'

GNAS1

McCune-Albright disease

174800

Acquire new substrate

PI (Pittsburgh allele)

1-Antitrypsin deficiency

107400

Ion channel inappropriately open

SCN4A

Paramyotonia congenita

168300

Structurally abnormal multimers

COL2A1

Osteogenesis imperfecta

Various

Protein aggregation

HD

Huntington disease

143100

Chimeric gene

BCR-ABL

Chronic myeloid leukemia

151410

Tumor CML Ewing sarcoma

Rearrangement t(9;22)(q34;q11) t(11;22)(q24;q12)

Chimeric gene BCR-ABL EWS-FLI1 EWS-ERG EWS-ATF1 EWS-WT1 FUS-CHOP FUS-ERG NTRK1-TPM3(TRK oncogene) E2A-PBX1 MLL-AFX1 MLL-AF4 MLL-AF9 MLL-ENL PML-RARA

Nature of chimeric product Tyrosine kinase Transcription factor Transcription factor Transcription factor Transcription factor Transcription factor Transcription factor Tyrosine kinase Transcription factor Transcription factor Transcription factor Transcription factor Transcription factor Transcription factor+retinoic acid receptor Transcription factor

Ewing sarcoma t(21;22)(q22;q12) (variant) ( ) Malignant melanoma of t(12;22)(q13;q12) soft parts Desmoplastic small round cell tumor Liposarcoma AML Papillary thyroid carcinoma Pre-B cell ALL ALL ALL ALL ALL Acute promyelocytic leukemia Alveolar rhabdomyosarcoma t(11;22)(p13;q12) t(12;16)(q13;p11) t(16;21)(p11;q22) inv(1)(q21;q31) t(1;19)(q23;p13.3) t(X;11)(q13;q23) T(4;11)(q21;q23) t(9;11)(q21;q23) t(11;19)(q23;p13) t(15;17)(q22;q12)

t(2;13)(q35;q14)

PAX3-FKHR

Rearreglos cromosmicos con produccin de genes quimricos

Gene PAX3 CFTR RET

Location 2q35 7p31.2 10q11.2

Diseases Waardenburg syndrome type 1 Alveolar rhabdomyosarcoma Cystic fibrosis Bilateral absence of vas deferens Multiple endocrine neoplasia type 2A Multiple endocrine neoplasia type 2B Medullary thyroid carcinoma Hirschsprung disease p g Charcot-Marie-Tooth neuropathy type 1A

Symbol WS1 RMS2 CF MEN2A MEN2B FMTC HSCR CMT1A HNPP PMC HYPP

MIM no. 193500 268220 219700 171400 162300 155420 142623 118220 162500 168300 170500

PMP22 SCN4A

17p11.2

Tomaculous neuropathy 17q23.1-q25.3 Paramyotonia congenita Hyperkalemic periodic paralysis Acetazolamide-responsive myotonia congenita 20p12-pter 20p12 pter 20q13.2 Xcen-q22 Creutzfeldt-Jakob Creutzfeldt Jakob disease Familial fatal insomnia Albright hereditary osteodystrophy McCune Albright syndrome McCune-Albright Testicular feminization syndrome Kennedy disease

PRNP GNAS1 AR

CJD FFI AHO PFD TFM SBMA

123400 176640 103580 174800 313700 313200

Gran heterogeneidad gentica CMT PMP22, peripheral myelin protein 22 MPZ m elin protein zero ero MPZ, myelin GJB1, connexin 32 EGR2, early growth response protein NDRG1, N-myc down regulated gene 1, MTMR2, myotubularin related protein 2 kinase GAN1, gigaxonin NEFL, neurofilament light chain KIF1B, kinesin 1B PRX, periaxin

Gene COL1A1

Location 17q22 17 22

Mutations Null ll l N ll alleles Partial deletions; substitutions N-terminal substitutions N terminal s bstit tions Deletion of exon 6

Syndrome OI t type I C-terminal OI type II

OI t pes I III or IV types I, EDS type VII OI type I

COL1A2

7q22.1

Splice mutations; exon deletions

C-terminal mutations N-terminal substitutions D l ti of exon 6 Deletion f COL2A1 12q13 Point mutations Nonsense mutation Defect i conversion f in i Missense

OI type II, IV OI type III EDS t type VII SED Stickler syndrome Kniest d l i i dysplasia Achondrogenesis II, spondylo-metaepiphyseal dysplasia

COL11A2

6p21.3

Splicing mutation

Stickler syndrome