Molecular cloning and expression of a pearl oyster (Pinctada fucata) homologue of mammalian putative tumor suppressor QM.
The QM gene was originally identified as a putative tumor suppressor gene from a Wilms' tumor cell line by subtractive hybridization assay. Later studies showed that the QM protein is multifunctional, involved in cell growth and differentiation, energy metabolism, respiration, and cytoskeletal function. In this report a full-length complementary DNA encoding a QM counterpart in pearl oyster (Pinctada fucata) was isolated. Phylogenetic analysis shows that oyster QM is more closely related to its insect homologues than to the mammalian homologues. Analysis of the tissue expression pattern of the oyster QM gene showed that oyster QM messenger RNA is expressed in all tissues tested, with highest levels in the digestive gland and mantle. Furthermore, we expressed the QM protein in Escherichia coli; Western blotting showed that the antibody of human QM is immunoreactive to the expressed oyster QM protein. Incubation of the oyster QM with Zn2+ resulted in the reduction of intrinsic emission fluorescence and a red-shift in the lambda(max) emission, indicating the occurrence of Zn(2+)-induced conformational changes. This evidence presents a possible mechanism for the critical function of zinc ion in the interaction of QM with Jun.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D010125:Ostreidae / Q000235:genetics*; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000080882:Ribosomal Protein L10; D012269:Ribosomal Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013050:Spectrometry, Fluorescence; D015032:Zinc
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; C000630686:RPL10 protein, human; D012269:Ribosomal Proteins; D015032:Zinc