Human papilloma viruses and cervical tumours: mapping of integration sites and analysis of adjacent cellular sequences.
BACKGROUND
In cervical tumours the integration of human papilloma viruses (HPV) transcripts often results in the generation of transcripts that consist of hybrids of viral and cellular sequences. Mapping data using a variety of techniques has demonstrated that HPV integration occurred without obvious specificity into human genome. However, these techniques could not demonstrate whether integration resulted in the generation of transcripts encoding viral or viral-cellular sequences. The aim of this work was to map the integration sites of HPV DNA and to analyse the adjacent cellular sequences.


METHODS
Amplification of the INTs was done by the APOT technique. The APOT products were sequenced according to standard protocols. The analysis of the sequences was performed using BLASTN program and public databases. To localise the INTs PCR-based screening of GeneBridge4-RH-panel was used.


RESULTS
Twelve cellular sequences adjacent to integrated HPV16 (INT markers) expressed in squamous cell cervical carcinomas were isolated. For 11 INT markers homologous human genomic sequences were readily identified and 9 of these showed significant homologies to known genes/ESTs. Using the known locations of homologous cDNAs and the RH-mapping techniques, mapping studies showed that the INTs are distributed among different human chromosomes for each tumour sample and are located in regions with the high levels of expression.


CONCLUSIONS
Integration of HPV genomes occurs into the different human chromosomes but into regions that contain highly transcribed genes. One interpretation of these studies is that integration of HPV occurs into decondensed regions, which are more accessible for integration of foreign DNA.
Klimov
Eugene
E
N,I, Vavilov Institute of General Genetics, RAS, Moscow, Russia. klimov_eugeney@mail.ru
Vinokourova
Svetlana
S
Moisjak
Elena
E
Rakhmanaliev
Elian
E
Kobseva
Vera
V
Laimins
Laimonis
L
Kisseljov
Fjodor
F
Sulimova
Galina
G
D001665:Binding Sites / Q000235:genetics; D002874:Chromosome Mapping; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D030561:Databases, Nucleic Acid; D005260:Female; D005819:Genetic Markers; D006801:Humans; D008969:Molecular Sequence Data; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000235:genetics / Q000821:virology; D021101:Radiation Hybrid Mapping; D017422:Sequence Analysis, DNA; D014412:Tumor Virus Infections / Q000235:genetics / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000821:virology*; D016662:Virus Integration / Q000235:genetics
D018076:DNA, Complementary; D005819:Genetic Markers