Zbornik - Društvo genetičara Srbije

II-Pos-4 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 59
Subotica, 30. novembar - 4. decembar 2004.
EKSPRESIJA I STRUKTURNA I FUNKCIONALNA ANALIZA
PROMOTORA HUMANOG SOX18 GENA
Isidora Petroviæ, Jelena Ðuroviæ, Gordana Nikèeviæ i Milena Stevanoviæ
Institut za molekularnu genetiku i genetièko inenjerstvo, Beograd
Familija SOX gena sadri veliki broj gena koji se eksprimiraju tokom embriogeneze, a
koji su u srodstvu sa sisarskim genom koji determiniše pol, oznaèenim kao SRY gen.
Iako je samo nekoliko SOX gena detaljno analizirano, sve dosadašnje studije ukazuju na
njihovu ulogu u regulaciji razviæa, a mutacije u nekim SOX genima su povezane sa
pojavom oboljenja kod èoveka.
Humani SOX18 gen je identifikovan pretraivanjem cDNK biblioteke napravljene iz
mozga humanog fetusa, korišæenjem specifiène SOXA probe. Mapiranjem pozicije gena
utvrðeno je da se SOX18 gen nalazi na hromozomu 20 u regionu q13,3. Pokazano je da
mutacije u SOX18 genu dovode do pojave recesivne i dominantne forme sindroma koji
obuhvata hipotrihozu, limfedem i teleangiektazije. Pored poznate funkcije u razvoju
dlake i krvnih sudova, ovo ukazuje i na vanu ulogu SOX18 transkripcionog faktora u
razvoju limfnih sudova.
U cilju odreðivanja molekularnih mehanizama odgovornih za kontrolu ekspresije
humanog SOX18 gena , analizirane su razlièite æelijske linije koje bi mogle da se koriste
kao model sistem za buduæe eksperimente. Ekspresija SOX18 gena je analizirana u
HUV-EC i EAhyb926 æelijskim linijama i detektovan je transkript duine 1,8 Kb.
Karakterizacija SOX18 promotora obuhvatala je odreðivanje starta transkripcije
metodom elongacije prajmera, kao i strukturne i funkcionalne analize regulatornog
regiona odgovornog za ekspresiju SOX18 gena. Analizirana je sposobnost razlièitih
fragmenata 5’ nekodirajuæeg regiona da utièu na ekspresiju cat reporter gena, a u cilju
identifikacije DNK regiona odgovornog za ekspresiju SOX18 gena.
EXPRESSION AND STRUCTURAL AND FUNCTIONAL ANALYSIS
OF THE PROMOTER OF HUMAN SOX18 GENE
The SOX gene family consists of a large number of embryionically expressed genes that
are related to the mammalian sex determining gene SRY. Although only few SOX genes
have been characterized in detail, all studies indicate that these genes have important
roles in the regulation of development and mutations in several SOX genes have been implicated
in human diseases.
Human SOX18 gene has been identified by screening human foetal cDNA library with
specific SOXA probe. Mapping analysis has revealed that SOX18 gene is located on human
chromosome 20q13.3. It has been shown that SOX18 mutations in human cause both
recessive and dominant hypotrichosis- limphedema and telangiectasia, suggesting that, in
addition to its established role in hair and blood vessel development, the SOX18 transcription
factor plays a role in the development and/or maintenance of lymphatic vessels.
In order to elucidate the molecular mechanisms controlling the expression of the human
SOX18 gene, we have analyzed different cell lines that could be used as a model system
for further experiments. The expression of SOX18 was studied by Northern blot analysis
in HUV-EC and EAhyb926 cell lines and a single transcript of about 1.8 kb was detected.
In order to characterize promoter of the SOX18 gene we have determined the transcription
start site by primer extension analysis and carried out structural and functional analysis
of the regulatory region responsible for SOX18 expression. To identify the DNA regions
responsible for the control of the SOX18 transcription, we have analyzed the ability
of truncated fragments from the 5’ flanking region to drive expression of cat reporter
gene.