Molecular cloning and expression analysis of xpd from zebrafish (Danio rerio). | - CCMAR -

Journal Article

TitleMolecular cloning and expression analysis of xpd from zebrafish (Danio rerio).
Publication TypeJournal Article
AuthorsSilva, IAL, M. Cancela, L, Conceição, N
Year of Publication2012
JournalMol Biol Rep
Volume39
Issue5
Date Published2012 May
Pagination5339-48
ISSN1573-4978
KeywordsAmino Acid Sequence, Animals, Cloning, Molecular, Conserved Sequence, DNA, Complementary, Gene Expression Profiling, Gene Expression Regulation, Developmental, Humans, Mice, Molecular Sequence Data, Open Reading Frames, Phylogeny, Protein Structure, Tertiary, Transcription Factor TFIIH, Xeroderma Pigmentosum Group D Protein, Zebrafish, Zebrafish Proteins
Abstract

The XPD gene, located in human chromosome 19, encodes one of the two helicase components of transcriptional factor IIH (TFIIH), a ten-subunit, multifunctional complex that is essential for multiple processes, including basal transcription initiation and DNA damage repair [1, 2]. Alterations in XPD resulting in defective TFIIH function are associated with UV-sensitive disorders including Xeroderma pigmentosum, Cockayne syndrome, and Trichothiodystrophy (TTD) [3, 4]. TTD mice exhibit many symptoms of premature aging, including osteoporosis, kyphosis and osteosclerosis [5]. This fact has triggered our interest in analyzing XPD involvement in bone biology using zebrafish as model organism. Although orthologs of xpd are present in all species analyzed, no specific data on its gene structure, regulation or function exists at this time in any fish system. In this study we isolated the zebrafish cDNA encoding xpd, and examined its spatial-temporal expression during early development as well as its tissue distribution in adult zebrafish. Only one gene was identified in zebrafish and its sequence analysis showed a molecular structure with 23 coding exons similar to other species. The amino acid sequences were also found to be largely conserved among all species analyzed, suggesting function maintenance throughout evolution. Gene expression analysis in different zebrafish tissues by qPCR showed xpd expression in all tissues examined with the highest expression in branchial arches. Analysis of xpd expression in zebrafish embryos showed maternal inheritance and presence of xpd transcripts in all developmental stages analyzed suggesting its implication in early zebrafish larval development.

DOI10.1007/s11033-011-1333-x
Sapientia

http://www.ncbi.nlm.nih.gov/pubmed/22187342?dopt=Abstract

Alternate JournalMol. Biol. Rep.
PubMed ID22187342
CCMAR Authors