丁金凤
中国医学科学院阜外医院 中心实验室
Myocyte's contraction is regulated by signal transduction pathway composed with many protein factors, but the definite mechanism is still uncertain. A novel cardiac-specific kinase gene which participates in regulation of signal transduction, p93 named, was cloned from adult heart cDNA library. p93, coding a family member of MAPKKK, localized on 1p31.1 based on bioinformatics analyses. Northern blot and 76-tissue array analyses determined that p93 was merely expressed in heart, but was undetectable in other tissues. Immunohistochemical study showed that p93 predominantly localized in the nucleus of cardiac myocytes. In vitro kinase assay indicated that p93 was a functional kinase capable of autophosphorylation. p93 could directly interact with cardiac troponin I by yeast two-hybrid system assessed utilizing bait plasmid containing p93 C-terminus (733 835 aa) and results were further confirmed by co-immunoprecipitation in vivo. Our data suggest that p93 is a cardiac-specific kinase and may play important role in regulation of sarcomeric contraction protein with signal transduction pathway similar ILK.
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica 2003
Cardiac-restricted genes play important roles in cardiovascular system. In an effort to identify such novel genes we identified a novel cardiac-specific kinase gene TNNI3K localized on 1p31.1 based on bioinformatics analyses. Sequence analysis suggested that TNNI3K is a distant family member of integrin-linked kinase. Northern blot and 76-tissue array analyses showed that TNNI3K is highly expressed in heart, but is undetectable in other tissues. Immunohistochemical analysis predominantly localized TNNI3K in the nucleus of cardiac myocytes. In vitro kinase assay showed that TNNI3K is a functional kinase. The yeast two-hybrid system showed that TNNI3K could directly interact with cardiac troponin I, results that were further confirmed by coimmunoprecipitation in vivo. Our data suggest that TNNI3K is a cardiac-specific kinase and play important roles in cardiac system.
Journal of molecular medicine (Berlin, Germany) 2003
OBJECTIVE:To focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (ASMC) by adeno-associated virus (AAV) vector carrying antisense thrombin receptor (ATR) and p21 double gene co-expression system.METHODS:Cultured human AMSC was infected with recombinant AAV containing ATR, p21 single gene and AP double gene respectively. The integration and expression of genes were confirmed by semi-quantitative RT-PCR. The anti-proliferation effect was determined by MTT assay. Cell cycle and apoptotic cell counts were measured through Flow Cytometry. The rate of apoptotic cells was examined with acridine orange/ethidium bromide(AO/EB) stain.RESULTS:RT-PCR indicated that the exogenous genes had been integrated into ASMC. The rates of cell survival were decreased by 16.67%, 21.60%, and 29.4% and the cell counts of G0/G1 phase were (61.8 +/- 2.9)%, (82.5 +/- 4.0)%, (80.4 +/- 6.1)% in ATR, p21 and AP group respectively after rAAV infected 4 days. The level and area of apoptotic peak were greater in AP double gene than ATR and p21 single gene. Cell stain indicated that apoptotic cells were (7.2 +/- 3.3)%, (10.7 +/- 5.6)%, and (18.3 +/- 2.7)% in each transgene group compared with (1.5 +/- 0.8)% in control group.CONCLUSION:AP double gene co-expression system has powerful effect for inhibiting proliferation and inducing apoptosis ASMC than ATR and p21 single gene and that is a superior way for gene therapy to restenosis.
Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae 2002
For searching cardiovascular-associated genes and investigating their expression profiles, human adult heart and aorta cDNA libraries were constructed, and a novel gene from adult heart cDNA library was isolated based on large-scale ESTs(expressed sequence tags) sequencing(GenBank accession number AF114264). The 2 736 bp clone contains one 1 344 bp open reading frame extending from 412 to 1 755. We named it NELIN (nexilin-like protein) because it shares high similarity with the rat nexilin. NELIN was expression-restricted in heart, skeletalmuscle, artery and vein by Northern blot and RT-PCR analyses, and mapped to chromosome 1p31-1p32 by database analyses. Based on domain structure, NELIN could regulate the formations of stress fibers,focal adhesion and its signaling complex, and even participates in the signal transduction in FAs(focal adhesions).
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica 2001
