Novel thyroid specific transcripts identified by SAGE: implication for congenital hypothyroidism - Color figures

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s

UvA-DARE (Digital Academic Repository)

Novel thyroid specific transcripts identified by SAGE: implication for congenital

hypothyroidism

Moreno Navarro, J.C.

Publication date

2003

Link to publication

Citation for published version (APA):

Moreno Navarro, J. C. (2003). Novel thyroid specific transcripts identified by SAGE:

implication for congenital hypothyroidism.

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W i f e J+ltittMtl

—— 1

PAXi!:.. NKX2.1 *

?? xlffl

11 1

Budding Budding ItógItóg ration SurvivalSurvival and Tifferentiation Tifferentiation Prenatal Prenatal expansion expansion FoHicis FoHicis formation formation G e n e e FKHL15 5 PAX8 8 NKX2.1 1 L o c u s s 9q22 2 2q13-14 4 14q12-q21 1 P r o t e i nn f u n c t i o n Transcriptionall regulation of geness involved in migration off the thyroid anlage and otherr midline developmental processes. .

Transcriptionall regulation of geness involved in survival andd differentiation of migrat-ingg thyroid cells.

P h e n o t y p e e

Agenesiss of the thyroid, cleftt palate and choanal atresia. .

Hypoplasiaa and/or ectopy off the thyroid, with severe CH. .

Choreoathetosis.. neona-tall respiratory distress andd thyroid hypoplasia and/orr TSH elevation.

Inher. .

A.R. . A.D. . A.D D O M I M M 602617 7 167415 5 600635 5

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Blood d

circulati i

TSI I

Gene e TSH-R R GNAS1 1 NIS S TG G TPO O PDS S THOX2 2 Locus s 14q31 1 20q13 3 19p13 3 8q24 4 2p25 5 7q31 7q31 15q21 1

--Proteinn function

Activationn of thyroid-specific meta-bolicc pathways.

Signall transduction from GPCRs for stimulationn of adenilyl cyclase Basall transport of iodine from blood streamm into the thyroid cell Matrixx (pro-hormone) for synthesis andd storage of thyroid hormones lodmationn of tyrosine residues of thy-roglobulinn (organification) and cou-plingg of iodotyrosines to form T3+T4. Transportt of iodide from the cytoplasm too the follicular lumen

Generationn of H202 in the thyroid

folli-cle. .

Dehalogenationn of MIT and DIT for iodinee recycling

Phenotype e

Thyroidd hypoplasia and severe CH Euthyroidd hyperthyrotropinemia

Resistancee to TSH and/or Albright's

heredi-tarytary osteodystrophy

Severee or moderate CH Euthyroidd goiter.

Goiterr and severe to moderate CH. Euthyroidd goiter.

Severee CH due to TIOD

"Pendred"Pendred syndrome" : deafness and goiter or

moderatee hypothyroidism due to PIOD. Permanentt and severe CH (TIOD) Transientt and moderate CH (PIOD). lodotyrosinee dehalogenase defect.

Inher. . A R . . A.R.A.D D A D . . AA R A R R A D D AR R A R R A R . . A.D. . A . R . A . D . . OMIM M 275200 0 103580 0 601843 3 600044 4 188450 0 274500 0 274600 0 607200 0 274800 0

ChapterChapter 1, fig. 4

££ Follicular

££ lumen

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CYTOPLASM M

Apical l

Membrane e

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Gene e HESX1 1 LHX3 3 PROP1 1 P 0 U 1 F 1 1 TRH H TRH-R R TSHA A TSHB B L o c u s s 3q21 1 9q3.4 4 5q q 3p11 ' 3q13 3 8q23 3 6q21 1 1p13 3 P r o t e i nn f u n c t i o n

Genee transcription regulation involved in differentiationn of all anterior pituitary cell lines and developmentt of optic nerve and brain. Genee transcription regulation invoked in differentiationn of all pituitary cell lines except for corticotropess and melanotropes

Genee transcnption involved in ontogenesis and differentiationn of all anterior pituitary cell lines, except thee corticotropes and melanotropes

Genee transcription involved in differentiation of thyrotropes.. lactotropes and somatotropes Activationn of TRH-receptor

Activationn of signal transduction leading to secretion off TSH in the thyrotropes

Activationn of GPCRs (dimehzation with chains). Activationn of TSH-receptor (dimenzation with TSH-a)

Phenotype e

Septo-oplicSepto-oplic dysplasia: optic nerve

hypoplasia.. CPHD and agenesis of midline brainn structures

Deficiencyy of TSH. GH. FSH and LH. and limitedd ability to rotate the head.

CPHD D

TSH.. GH and PRL deficiencies.

Nott reported

Isolatedd central hypothyroidism of late onsett and growth retardation Glycoproteinn hormones deficiency. Isolatedd hereditary TSH deficiency.

Inher. . AA R AA R AA R AA R AA D AA R AA R OMIM M 601802 2 600577 7 601538 8 173110 0 275120 0 188545 5 118850 0 188540 0

ChapterChapter 1, fig. 6

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Gene e TRa a TRB B THH transporters DI01 1 DI02 2 DI03 3 Locus s 17q11 1 3p23 3 1p32 2 14q24 4 14q32 2 Proteinn function

Recruitmentt of nuclear T3 and transcriptionn of T3 target genes. Recruitmentt of nuclear T3 and transcriptionn of T3 target genes. Transportt of T4/T3 into peripheral

cells. .

Deiodinationn of T4 into T3 in liver andd kidney.

Deiodinationn of T4 into T3 in pituitary,, thyroid, muscle and brain.

Deiodinationn of T3 into reverse T3 orr T2 in placenta, brain and vessels. .

Phenotype e

Nott identified in human.

Unresponsivenesss to thyroid hormone. .

Nott described.

Euthyroidd hyperthyroxinemia Mutationss not identified in humans.

Nott described (Pituitary TH resistance,, suspected).

Severee hypothyroidism in giant hemangiomas.. No mutations identified. . Inher. . Not t reported d A.R,, A D .

--Not t reported d Not t reported d Not t reported d OMIM M 188450 0 190160 0

--147892 2 601038 8 601413 3

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•AAAAA A

Isolatee SAGE tags

Linkk tags together

Sequencee linked tags

Quantitatee tags and determine

patternss of gene expression

-n.Jlnll l

AA a C TI r r £ v

Genee product

N o r m a l l

AA T» f. TI V t

Genee product

Disease e

ChapterChapter 1, fig. 8

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tTTTT--Cleavee w th anchoring enzyme iAE)

Bindd to streptavidm beads

Dividee in ^

Ligctee to linkers IA + B)

II.TM M

II ,rv

I I

Cleavee with tagging enzyme fTE)

Bll .

XXXXXXXXX X

G1ACXXXXXXXXX X

i i

TEE AE Tag

lOOOOOOOOO O

cc TACCTACOCOOOOOOO

TEE AE

Tag g

V AA Ugate ano amplify with ^

" ^^ primers A and B ^

GGAA XXXXXXXXXOOCOCXXXXtCATGCATC

G^ACXXXXXXXXJCCKXOOOCOOGTACCT T

** Ditaq

r

I I

Cleavee with anchoring enzyme

Isolatee ditags

Concatenatee and clone

-CATT £XXXXXXXXXOOOCXX>COC^

GTT ACXXJQ0CXXXXCK>OCXX5O0OGTACX^^

-"-- lag J ? - " - ^ W a g 3—> W a g 4—^

AEE

cc V

"

T a g 1

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Mutants s

Controls s

5 '' A C G G G G Ü V 5 ' A C G G G I I G A G A

: :

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**'LL 'LL**

--C1300T T

5'' Y & A G 6 A r, y 5 A A r A A i. i, i, (, y

V V

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C2101T T

5"" G G G i Y '• G G C A 3 ' s_{' G G G G 0}

üü

/11 A

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C2056T T

< — > >

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^

_A _A

2895-2898del l

ChapterChapter 3, fig. 1

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T H O X 2 2 p r o t e i nn 2 , R434X X S965fsX994 4 Q686XX R701X -COOH H O — OO 50 amino acids C Z I I C=> > H3 H3 H i i Transmembranee domain EF-nanrJJ motif FAD-bindingg motif NADPH-bindingg motif Putativee glycosylation site Putativee heme-binding site

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a a

E x o n s s 85.433 3 1 1 ATG G 56.090 0 22 3 4 5 6 TGAA TAG

++ -I-+4

alternative e splicing g Genomicc clone AL031010 0 Genome e (Chr.. 6q24) DEHAL1.A DEHAL1.A (ORF:: 867 bp) DEHAL1.B DEHAL1.B (ORFF 876 bp

ChapterChapter 4, fig. 1

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H s D E H A L I AA 289 aa H s O E H A M BB 292 aa MmMm NP_081667 285 aa DmAAM110099 287 aa DmDm NP_64843 757 aa ScNP_6294666 226 aa PfPfNP_579385NP_579385 187 aa TtTt Q60049 205 aa Ecc P38489 217 aa EcP171177 240 aa II Signal anchor II N i t r o r e d u c t a s e d o m a i n HsHs D E H A L 1 A H ss D E H A L 1 B MmMm NP_081667 DmDm A A M 1 1 0 0 9 DmDm NP_64843 ScSc NP_629466 PfPf NP_579385 TtTt Q60049 EcEc P38489 EcEc P17117 E|FF Y E L L N K | 3 R B V [ 3 F I - S N E Q V PJM E V E F Y E L L N K R R S V P . F II - S N E Q V P M E V E F Y E L L N K R R S V R F I - S S E H V P M E V V R L Y E L M R G R R S I R S F S S H P K P D L S V V R L Y E L M R G R R S I R S F N S H P K P D L S V V DD N V I R T A G T A U r - f J A H I T E P W T F V V ~ V ] K D P D N VV I R T A G T A P S G A H T E P W T F V V V K D P E N V I K A A G T A P S G A H T E P W T F V V V K D P P E D C I R A A G T A P S G A H T E P W T Y C V V Q E P P E D CC I R A A G T A P S G A H T E P W T Y C V V Q E P D F V S F V D R R R S V R F F - S D E P V P R E C L D L A V A A A N T A P S G A H F Q P W K F A V I G D A A R V L E L A K R R K T V R K FF - S K D R P P I E L V F K A I E T A K E A P S G M N A Q P W K F V V I D N P RR D P D [ A KK T A A L K H R H I H R Y - R K D P DD I I S V A L K y H C j T Q A F D A S K K PP T I E L I C G H R B I 0 H F - T D E P VV P I E G L L R E I L E A A | L R A P S | A W N L Q P W R I V V V LL T i P E Q A E Q I K T L L Q Y S p | II S E A Q R E A I I N S A R | A T | SS Q P W H F I V A L Q C S S II I R I

ss

T D K K T £ Hss D E H A L 1 A HsHs D E H A L 1 B MmMm NP_081667 DmDm A A M 1 1 0 0 9 DmDm NP_64843 ScSc NP_629466 PfPf NP_579385 TtTt Q60049 EcEc P38489 EcEc P17117 H ss D E H A L 1 A H ss D E H A L 1 B MmMm NP_081667 DmDm A A M 1 1 0 0 9 DmDm NP_64843 Scc NP_629466 PfPf NP_579385 TtTt Q60049 EcEc P38489 EcEc P17117 DD V K H K DD V K H K D M K H K K EE L K R S EE L K R S AA T K H R EE L K A R RR K RR K RR E RR F RR E II E E E E E I N Y M K R | M G - H i R W V T D L K K L R T N W I K E Y L D T A P I L I L I F II E E E E E I N Y M K R M G - H R W V T D L K K L R T N W I K E Y L D T A P I L I L I F II E E E E E I N Y M K R M G - K R W V T D L K K L R T N W I K E Y L D T A P V L I L I F V E Q E E L V N Y S Q R M HH - P Q W V T D L R P L Q T N H V K E Y L T E A P Y L I L I F V E Q E E L V N Y S Q R M HH - P Q W V T D L R P L Q T N H V K E Y L T E A P Y L I L I F R V A A E E E E R V N Y E G G R I P P E W R A A L A R L E T D S D K G F L D V V P W I V V C F F R E V C E V E E E K F Y E RR I K G - - E L K E W L V E N E F T P E K P F L T E A P Y L V L V F AA T K R A L R E A|A - - - - • F g ] E G K A R V A K S A A G N Y Y A L R E E L V U ll T G G K~Q~|vv H G F A A N G K K K V H Y Y N E I S V S I A C G I L L A A L Q N A G L V T V IMMtp L K Q V H G F AA A N G K K K V H Y Y N E I S V S I A C G I L L A A L Q V N N G I T M R t Q I A R K Q V H G F AA A N G K K K V H Y Y N E I S V S I A C G L L L A A L Q N A G L V T V T T T P L K Q T Y G L SS E N G K R M R R H Y Y N E I S T S I A A G I L L C A L Q A A G L A S L V T T P L K Q T Y G L SS E N G K R M R R H Y Y N E I S T S I A A G I L L C A L Q A A G L A S L V T T P L A E K S T A LL P D G S L R K N Y Y V N E S V G I A C G L F I T A L H A M G L S T L T H T P GG Y • T K A P Y W L Q S V W I A V G Y F L L A L E E V E L G T V T Y T P -80|LL E D A L A|H L [24] A A M G Q E A R K A W A S G Q S Y I L L G Y L L L L L E AJ Y G L G S V B AA K T A M D P VWLl[32] H R K D L H D D A E W M A K Q V J ^ J N V G N F L L G V AIA L G L : P A | V P AA D F N R H L Q l~[c P D A Q L G L A E Q L L L G vflD T A M M A Q N A L I A A E [ S L G L G G V| Hss D E H A L 1 A Hss D E H A L 1 B MmMm NP_081667 DmDm A A M 1 1 0 0 9 DmDm NP_64843 ScSc NP_629466 PfPf NP_579385 TtTt Q60049 EcEc P38489 EcEc P17117 NN C G P R L R V L L|G R P - A H[E K L L E L L P V G ] Y P S K E - A T V P D L K R Q P L D Q I M|V T V[ H R H LL I E G P G R S S E A C S K L S S Q G R P E C R S G D C H Y H S S Q L W P S T E G A P G P P R N C G P R L R V L L G R P - S H E K L L V L L P V G Y P S R D - A T V P D L K R K A L D Q I M V T V V N C G P A L R N L L G R PP - V N E K L L I L L P V G Y P K D G - C T V P D L A R K N L S N I M V T F N C G P A L R N L L G R P - V N E K L L I L L P V G Y P K D G - C T V P D L A R K N L S N I M V T F F N P M A F L T RR I C A R P - G N E R P Y I L F P V G Y A A P D - C E V P D L A R K P L A E A I T E P PP N T R P I E E L L Q V P - R E W R L O V I L P V G Y P D D P - - K P K L K R K S I E E V V S F N FF D P E R V R A I L G L P S RIA A I P A L V A L Gl Y P A E F D A A I L D A E F G L K E K G G NNIII E A V T K L L K L P - Q H A AA I P A L V A L G Y T S L V V V P V G G V L P L F G L C L G G -- - - E G Y P S H HH H S V E DJF N A[ T L P Q S Q I W PP A D -L P -L E R V V -L W W

I I

-- • N P D L l a P Q L P A S I L LL T E VV H E S e c o n d a r yy s t r u c t u r e FMNN interaction

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B-- ..

2 2

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21,81921,819 bp _{Genomicc clone} AC007011 AC007011

II I I II

E x o n ss 1 2 3 4 5 6 88 1 kb G e n o m e e (Chr.. 16p) aacc aacc Arc c

--I --I t t t t • •

--. --.

N M 4 11 c R N A 1.3451.345 bp ORF:ORF: 1.083 bp N M 4 11 p r o t e i n 361361 ammo acids