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A Novel Truncated Form of DNA Polymerase ß Protein in Mouse and Zebrafish Early Stage Embryos
Author(s) -
Dobosh* Brian,
Kaplan* Nicole,
Moore Stephen,
Strauss Phyllis
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.879.17
Subject(s) - biology , zebrafish , microbiology and biotechnology , embryo , polyclonal antibodies , blot , gene , antibody , genetics
This laboratory has been studying base excision repair (BER) in early embryogenesis using zebrafish as a model system. We originally reported that unfertilized zebrafish eggs and early stage embryos lack DNA polymerase beta protein ( Polβ) despite ample amounts of polb mRNA. Here we report that early stage zebrafish embryos and pre‐implantation mouse oocytes and embryos do contain Polβ. However, the major form is a truncated 26 kDa protein detected using 18S monoclonal antibody directed against amino acids 140‐154 in mouse samples and a custom rabbit polyclonal antibody directed against the last few residues of the zebrafish carboxyl terminus by Western blotting. When the Western blots are probed using an antibody designed to detect the 8kDa amino terminus, no signal is detected. The same truncated protein is also found in mouse ovarian follicle cumulus cells in addition to the 39 kDa canonical form. The truncated form is reflected in qRT‐PCR results that show enhanced transcription of the 3' terminus of the gene which corresponds to the previously approved expressed sequence tags (ESTs) EG587345 in zebrafish, BY759906 in the amniotic fluid of pregnant mice, and CJ228856 in mouse embryos. We conclude that a novel form of Polβ protein is present in unfertilized oocytes and early stage embryos. (Supported with funds from Aid for Cancer Research and the Provost's Fund to Support Undergraduate Research, Northeastern University) *These two students contributed equally to this body of work.