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          Institute: MPI für molekulare Genetik     Collection: Research Group Development and Disease     Display Documents

ID: 541483.0, MPI für molekulare Genetik / Research Group Development and Disease
Analysis of relative gene dosage and expression differences of the paralogs RABL2A and RABL2B by Pyrosequencing.
Authors:Kramer, M.; Backhaus, O.; Rosenstiel, P.; Horn, D.; Klopocki, E.; Birkenmeier, G.; Schreiber, S.; Platzer, M.; Hampe, J.; Huse, K.
Research Context:This work was supported by grants from the National Genome Research Network (NGFN 01GS0121, 01GS0809 to S.S. and M.P.) and the Deutsche Forschungsgemeinschaft (DFG Hu498/3 to K.H. and Ha3091/2-1 to J.H.).
Date of Publication (YYYY-MM-DD):2010-05-01
Title of Journal:Gene
Journal Abbrev.:Gene
Issue / Number:1-2
Start Page:1
End Page:7
Copyright:© 2010 Elsevier B.V. All rights reserved.
Review Status:not specified
Audience:Experts Only
Abstract / Description:The paralogous genes RABL2A (chr2) and RABL2B (chr22) emerged by duplication of a single gene in the human-chimpanzee ancestor and share a high degree of sequence similarity. In Phelan-McDermid-Syndrome microdeletions of 22q13 often also affecting RABL2B are of clinical importance but their incidence is still unknown. We analyzed a German population (190 individuals) for such aneuploidies and the paralogs' expression in cell lines by RABL2 paralogous sequence quantification. For determination of the genomic and transcriptional ratios of RABL2A and RABL2B a Pyrosequencing protocol was introduced as a high-throughput method. During PCR the 3' end of the biotinylated strand is engineered by a backfolding oligonucleotide to hybridize in the Pyrosequencing reaction to an internal site near the sequence to be analyzed. In human samples no deviations of the euploid genomic state could be detected indicating that 22q13 microdeletions involving RABL2B are rare. However, despite equal gene dosage a preferential expression of RABL2B in human tissues and lymphoblastoid cell lines was detected which is most pronounced in brain and placenta. This renders a complete functional complementation of one paralog by the respective other unlikely and hints to a functional and clinical importance, in particular with respect to the 22q13 chromosomal deletion syndrome. Remarkably and in contrast to human, expression levels of the two paralogs in a chimpanzee cell line are equal. This finding is discussed in view of the relocation of RABL2A from its ancestral telomeric to its pericentromeric location in human.
Free Keywords:Paralogs;
Gene expression;
Paralog ratio test;
Comment of the Author/Creator:Corresponding author: Klaus Huse
Tel.: + 49 3641 656254; fax: + 49 3641 656255.
External Publication Status:published
Document Type:Article
Communicated by:Stefan Mundlos
Affiliations:MPI für molekulare Genetik
External Affiliations:1.Leibniz Institute for Age Research—Fritz Lipmann Institute, Genome Analysis, Beutenbergstr. 11, 07745 Jena, Germany;
2.Institute of Clinical Molecular Biology, University Hospital Schleswig-Holstein, Christian Albrechts University, Kiel, Germany;
3.Institute of Medical Genetics, Charite, University Medicine of Berlin, Germany;
4.Institute of Biochemistry, Medical Faculty, University of Leipzig, Leipzig, Germany;
5.Department of Internal Medicine I, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Identifiers:ISSN:0378-1119 [ID No:1]
URL:http://www.ncbi.nlm.nih.gov/pubmed/20138207 [ID No:2]
DOI:10.1016/j.gene.2010.01.005 [ID No:3]
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