Biomedical subjects

Y Ye

Publications and source records attributed to Y Ye.

At least 73 records · Page 4Linked to original sources

PubMed2000

[AP-1 mediated signal transduction in thrombin-induced regulation of PAL-1 expression in human mesangial cells].

OBJECTIVE: To evaluate activator protein-1 (AP-1) mediated mechanisms in thrombin-induced qlasminogen activator inhibitor-1 (PAI-1) expression in cultured human glomerular mesangial cells (MCs). METHODS: Electrophoretic mobility shift assay (EMSA) was employed to assess AP-1 DNA-binding activity, and Western blot hybridization was used for quantification of c-fos and c-jun, two subunits of AP-1 dimers. PAI-1 activity and mRNA expression were analysed by the fibrin plate assay and Northern hybridization, respectively. RESULTS: Thrombin concentration enhanced PAI-1 activity in the supernatant and stimulated PAI-1 mRNA expression in cultured MCs. PAI-1 activity was blocked by hirudin, a specific inhibitor of thrombin. Further study demonstrated that thrombin promoted AP-1 DNA-binding activity but exerted little effect on c-fos or c-jun. Curcumin (AP-1 inhibitor), staurosporine (PKC inhibitor), and genistein (PTK inhibitor) all reduced AP-1-mediated PAI-1 mRNA expression induced by thrombin in cultured MCs. CONCLUSION: The present study indicates that in cultured human MCs, thrombin stimulates PAI-1 expression through an AP-1 signal pathway, which may be mediated by PKC and PTK.

Cells, Cultured↗

PubMed2000

[Effect of hormone replacement therapy on coagulation function in postmenopausal women].

OBJECTIVE: To study the effect of hormone replacement therapy (HRT) on coagulation function in postmenopausal women. METHODS: Fifty-two postmenopausal women were divided into three groups; placebo group (10 cases), estrogen group (19 cases) and estrogen plus progesterone group (23 cases). Twenty healthy premenopausal women served as control group. The coagulation parameters were determined before and 3, 6 months after initiation of HRT. RESULTS: The levels of fibrinogen and protein S in postmenopausal women were significantly higher than those in the control group. There was no difference in coagulation parameters before and after placebo administration. After 3-month HRT, the fibrinogen levels in estrogen group and estrogen plus progesterone group significantly decreased and further decreased after 6-month HRT, as compared with before HRT. A significant decrease was also noted in protein S in both groups after 3 month HRT, but returned closely to baseline after 6-month HRT. There was no significant difference in coagulation parameters between estrogen group and estrogen plus progesterone group either before or after HRT. CONCLUSION: HRT significantly decrease the levels of fibrinogen in postmenopausal women.

Blood Coagulation↗

PubMed2000

[Study on the amplification uniformity of whole genome in a single cell by degenerate oligonucleotide primed polymerase chain reaction].

OBJECTIVE: To investigate the uniformity of whole genome amplification from a single cell by degenerate oligonucleotide primed polymer ase chain reaction (DOP-PCR). METHODS: Amplification of the whole genomic DNA from a single cell with XX, XY, XO, XXY, +13 or +21 was performed by DOP-PCR. Comparative genomic hybridization (CGH) of a single cell DOP-PCR product against the genomic DNA or a single cell DOP-PCR product from normal male as carried out respectively. RESULTS: The average profile of fluorescence intensive ratio in CGH with the genomic DNA as the reference was greatly fluctuated and the standard deviation in about 30% haploid was beyond the normal limits. False positive hyperrepresentation was existed in X chromosome while trisomy 13 and 21 were failed to be detected. However, the distributions of the mean and the standard deviation of the ratio in the CGH with DOP-PCR product as the reference were acceptable. The copy number changes of chromosome X, Y, 13 and 21 were demonstrated. CONCLUSION: Although the single cell DOP-PCR product is not completely uniform, it would be reliable to study the whole genome of a single cell by controlling the conditions of PCR and selecting the suitable control reference because the unequal amplification is unrandom.

Female↗

PubMed2000

[Relationship between tumor suppressor gene p16 and Rb and early diagnosis of lung cancers].

OBJECTIVES: To study the incidences and mechanisms of tumor suppressor gene p16 and Rb inactivation in lung cancers, and to investigate their relation to biological characters, clinical pathological diagnosis and gene classification diagnosis of lung cancers. METHODS: Immunohistochemistry, double in situ hybridization, PCR, PCR-SSCP and sequencing were applied to study the changes of p16 and Rb genes in 106 lung cancers and 23 lung tissues of benign diseases. RESULTS: Total expression rates of p16 and Rb proteins and mRNAs in lung cancers were lower than those in normal lung tissues and benign lung diseases, and they were specially related to histological type, metastasis and clinical stage of lung cancers. In stage I and II lung cancers, the obvious inactivation of tumor suppressor gene p16 or Rb was examined (32.6% or 28.3%); p16 inactivation was detected mainly in non-small cell lung cancers, and Rb inactivation mainly in small cell lung cancers. There were three mechanisms of homozygous deletions, methylations and mutations for p16 gene inactivation. The deletion rate of p16 exon1 and/or exon2 was 25.8%, mainly took place in p16 protein negative cases of non-small cell lung cancers. 15 cases (16.9%) took part in methylations on SmaI sites of CpG island of p16 gene. PCR-SSCP and sequencing showed that 9 cases had p16 gene mutations. CONCLUSIONS: p16 and Rb genes may play important roles in genesis and progression of lung cancers. Inactivation of p16 or Rb gene may be is an early link of lung canceration, that is very important for early diagnosis of lung cancers. A new gene classification model for lung cancer diagnosis would be set up based on the research of p16 and Rb genes.

Adolescent↗

PubMed2000

[CDKN2/p16 gene cloning and its probe preparation and application in lung cancer].

OBJECTIVES: To clone CDKN2/p16(INK4a) gene, prepare its probe, and to study the change of CDKN2/p16(INK4a) gene in lung cancers. METHODS: Total RNA of normal lung tissue was extracted, CDKN2/p16(INK4a) gene cDNA synthesized, and CDKN2/p16(INK4a) gene recombinant vector, constructed. Southern blot was used to study CDKN2/p16(INK4a) gene in 46 cases of lung cancers, 3 cases of normal lung tissues, 6 cases of lung tissues near cancer, and 3 cases of lymph nodes with lung cancer metastasis. RESULTS: Cloned CDKN2/p16(INK4a) cDNA was proved by enzyme digestion and sequencing. Southern blot showed 4.3 kb band in normal lung tissues and lung tissues near cancers, and deletion of CDKN2/p16(INK4a) gene in cancer tissues and lymph nodes with lung cancer metastasis, with a deletion rate of 17.4% (8/46). CONCLUSION: CDKN2/p16(INK4a) gene may play a role to some extent in progression of lung cancers.

Blotting, Southern↗

PubMed2000

Relations of tumor suppressor gene p16 and RB to early diagnosis of lung cancer.

OBJECTIVES: To study the incidence and mechanism of the tumor suppressor gene p16 and RB inactivation in lung cancer, and to investigate their relations to biological characteristics, clinical pathological diagnosis and gene classification. METHODS: Immunohistochemistry, double in situ hybridization, PCR, PCR-SSCP, and sequencing were used to study the changes in the p16 and RB genes in 106 patients with lung cancer. The specimen of normal lung tissue from the patients with lung cancer and specimens of the 23 patients with benign lung diseases were studied in contrast to those of the same patients. RESULTS: The total expression rate of the p16 and RB genes in the lung cancer tissues of the 106 patients was lower than that in normal lung tissues and tissues of benign lung diseases and was specially related to histological type, metastasis, and clinical stage of lung cancer. In stage I and II lung cancer, the obvious inactivation rate of the tumor suppressor gene p16 or RB was 32.6% or 28.3%. The p16 inactivation appeared largely in non-small cell lung cancer (50.1%), and the RB inactivation largely in small cell lung cancer (88.2%). Homozygous deletion, methylation, and mutation were three mechanisms for the p16 gene inactivation. The deletion rate of the p16 exon 1 and/or exon 2 was 25.8% in the patients with non-small cell lung cancer. Fifteen patients (16.9%) showed methylation at the SmaI sites of CpG island of the p16 gene. PCR-SSCP and sequencing revealed p16 gene mutation in 9 of the 106 patients with lung cancer. CONCLUSIONS: The p16 and RB genes may play an important role in genesis and progression of lung cancer. The inactivation of the p16 or RB gene may be an early sign of lung canceration, that is important for the early diagnosis. A new gene classification model for lung cancer diagnosis would be set up based on the research of the p16 and RB genes.

DNA Methylation↗

PubMed2000

[Hyperpolarized cardiac arrest with ATP-sensitive potassium channel opener on myocardial protection during CPB].

OBJECTIVE: To investigate the myocardial protective effects of pinacidil-induced hyperpolarized arrest and compare them with those induced with depolarized hyperkalemic arrest. METHODS: 18 dogs were equally divided into three groups. In the hypothermic hyperpolarization group (LH group), after aortic cross-clamping, a single dose of 4 degrees C pinacidil containing St. Thomas cardioplegic was infused through the aortic root. Temperature during CPB was kept between 26 - 28 degrees C and warmed to 37 degrees C before aortic declamping. Global ischemia lasted 60 min and then reperfusion started for 30 min. In the normothermic hyperpolarized group (WH group), the same procedure was set as in the LH group, except maintaining temperature of 35 - 37 degrees C for CPB and pinacidil solution. In the control group (group C), no pinacidil in St. Thomas solution was the only difference to the other 2 groups. Cardiac arrest and its recovery, the ultrastructure of the myocardium and hemodynamic during ischemia and after reperfusion were observed in the 3 groups. RESULTS: (1) The percentages of normal mitochondria and glycogen were not changed significantly during ischemia and after reperfusion in the LH group, but declined markedly in the group C at ischemic 30, 60 min, and reperfusion for 20 min (P < 0.01). In the WH group, they were lower than those of the group LH, but higher than those of the group C before ischemia. (2) The recoveries of CO, SV, CI, LVSW, RVSW, MAP in the LH group were significantly better than those in the other two groups after reperfusion for 15 minutes and 30 minutes (P < 0.05 or 0.01). However, they were much better in the WH group than in the group C (P < 0.05 or 0.01). (3) The time from cardioplegic infusion to cardiac arrest was shorter in the group C and the group LH than in the group WH. CONCLUSIONS: Myocardial protection for global ischemia during CPB could be well achieved with hyperpolarized cardiac arrest induced by ATP-sensitive potassium channel opener, pinacidil, especially in the hypothermic state. The protection is weaker in normothermia but is still stronger than that with traditional depolarized arrest.

Adenosine Triphosphate↗

PubMed2000

[Study on mechanism of Shenle and Fragmin in preventing and treating glomerulosclerosis in rats].

OBJECTIVE: To study the effects of Shenle and Fragmin on the degradation of extracellular matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-1 (TIMP-1) in the remnant kidneys of 5/6 nephrectomized rats treated with Shenle and Fragmin. METHODS: The model of focal and segmental glomerulosclerosis was made by 5/6 nephrectomy in Wistar rats. After being treated with Shenle (4 g.kg-1.d-1) or Fragmin (1000 IU.kg-1.d-1) for 20 weeks, the blood pressure, proteinuria, serum creatinine, lipids, renal pathological change of remnant kidney of the model animals were examined, and the mRNA expression of MMP-9, TIMP-1 and type IV collagen in the remnant kidney were also detected. RESULTS: Shenle and Fragmin could significantly reduce the blood pressure, proteinuria, serum creatinine and lipids, alleviate the pathologic change of kidney tissue, decrease the type IV collagen, MMP-9 deposition. Shenle and Fragmin down-regulated the mRNA expression of MMP-9 by 30.0% and 28.5% and TIMP-1 expression by 59.3% and 55.0% in the remnant kidney of the 5/6 nephrectomized rats respectively. CONCLUSION: Shenle and Fragmin might ameliorate glomerulosclerosis through modulating MMP-9/TIMP-1 gene expression, alleviate the abnormal metabolism and accumulation, promote extracellular matrix degradation in glomeruli remodeling, it might be the important mechanism of Shenle and Fragmin in treating glomerulosclerosis.

Animals↗

PubMed2000

[Numerical taxonomy of medicinal plants from Areae in Zhejiang Province].

Numerical taxonomic studies were done on 13 species from 3 genera of tribe Areae in Zhejiang, used as 13 operational taxonomic units (OTUS). 40 morphological characters were used for analysis. Euclid distance coefficients used to show quantitative index of similarity among OTUS were computed by standardized data. The dendrograms from 8 systematic cluster methods including single linkage method, complete linkage method, median method (beta = 0 WPGMA) and (beta = -0.25 WPGMA), centroid method, group average method, variable group average method (beta = -0.25) and variable method (beta = 0.25) were constructed respectively. WPGMA was slected as the optimal one by computing the comparative coefficients of every cluster result and the boundary of taxa in its dendrogram determined by a method to treated IBM computer with the program by BASIC language. The Results showed that 13 OTUs were classified as 3 clusters by broken line L2, namely, cluster I: Typhonium; cluster II: Arisaema; cluster III: Pinellia, and cluster II further as 4 groups by L1, which belong to Section Pistillata, Arisaema, Sinarisaema and Tortuosa respectively. Being consistent with those from classical taxonomy, the results in this study may be helpful to the classification and identification of the medicinal plants from tribe Areae in Zhejiang Privince.

Araceae↗

PubMed1999

Characterization of two chitinase genes and one chitosanase gene encoded by Chlorella virus PBCV-1.

Chlorella virus PBCV-1 encodes two putative chitinase genes, a181/182r and a260r, and one chitosanase gene, a292l. The three genes were cloned and expressed in Escherichia coli. The recombinant A181/182R protein has endochitinase activity, recombinant A260R has both endochitinase and exochitinase activities, and recombinant A292L has chitosanase activity. Transcription of a181/182r, a260r, and a292l genes begins at 30, 60, and 60 min p.i., respectively; transcription of all three genes continues until the cells lyse. A181/182R, A260R, and A292L proteins are first detected by Western blots at 60, 90, and 120 min p.i., respectively. Therefore, a181/182r is an early gene and a260r and a292l are late genes. All three genes are widespread in chlorella viruses. Phylogenetic analyses indicate that the ancestral condition of the a181/182r gene arose from the most recent common ancestor of a gene found in tobacco, whereas the genealogical position of the a260r gene could not be unambiguously resolved.

Amino Acid Sequence↗

PubMed1999

Apoptotic activities of wild-type and Alzheimer's disease-related mutant presenilins in Drosophila melanogaster.

Mutant human presenilins cause early-onset familial Alzheimer's disease and render cells susceptible to apoptosis in cultured cell models. We show that loss of presenilin function in Drosophila melanogaster increases levels of apoptosis in developing tissues. Moreover, overexpression of presenilin causes apoptotic and neurogenic phenotypes resembling those of Presenilin loss-of-function mutants, suggesting that presenilin exerts a dominant negative effect when expressed at high levels. In Drosophila S2 cells, Psn overexpression leads to reduced Notch receptor synthesis affecting levels of the intact approximately 300-kD precursor and its approximately 120-kD processed COOH-terminal derivatives. Presenilin-induced apoptosis is cell autonomous and can be blocked by constitutive Notch activation, suggesting that the increased cell death is due to a developmental mechanism that eliminates improperly specified cell types. We describe a genetic model in which the apoptotic activities of wild-type and mutant presenilins can be assessed, and we find that Alzheimer's disease-linked mutant presenilins are less effective at inducing apoptosis than wild-type presenilin.

Alleles↗

PubMed1999

Neurogenic phenotypes and altered Notch processing in Drosophila Presenilin mutants.

Presenilin proteins have been implicated both in developmental signalling by the cell-surface protein Notch and in the pathogenesis of Alzheimer's disease. Loss of presenilin function leads to Notch/lin-12-like mutant phenotypes in Caenorhabditis elegans and to reduced Notch1 expression in the mouse paraxial mesoderm. In humans, presenilins that are associated with Alzheimer's disease stimulate overproduction of the neurotoxic 42-amino-acid beta-amyloid derivative (Abeta42) of the amyloid-precursor protein APP. Here we describe loss-of-function mutations in the Drosophila Presenilin gene that cause lethal Notch-like phenotypes such as maternal neurogenic effects during embryogenesis, loss of lateral inhibition within proneural cell clusters, and absence of wing margin formation. We show that presenilin is required for the normal proteolytic production of carboxy-terminal Notch fragments that are needed for receptor maturation and signalling, and that genetically it acts upstream of both the membrane-bound form and the activated nuclear form of Notch. Our findings provide evidence for the existence of distinct processing sites or modifications in the extracellular domain of Notch. They also link the role of presenilin in Notch signalling to its effect on amyloid production in Alzheimer's disease.

Alzheimer Disease↗

PubMed1999

In vitro and in vivo studies of 1H NMR visibility to detect deoxyhemoglobin and deoxymyoglobin signals in myocardium.

1H nuclear magnetic resonance (NMR) spectroscopy can be used noninvasively to detect the proximal histidyl N delta proton signals of deoxymyoglobin in the myocardium. However, the quantification of deoxymyoglobin is based on the assumption that the deoxymyoglobin signal detected is not contaminated by the deoxyhemoglobin signals contributed from the blood. The purpose of this study was to conduct in vitro and in vivo 1H NMR studies to examine the in vivo NMR visibility of deoxyhemoglobin in the myocardium. The results demonstrate that the NMR visibility of alpha and beta subunits of deoxyhemoglobin is sensitive to the pulse width for spin excitation because of short T2 relaxation times, and they are not NMR visible in the canine myocardium in vivo at 4.7 T when a 0.5-1.0 msec long Gaussian excitation pulse is used. Therefore, the resonance peak detected at approximately 72 ppm (relative to the water resonance) in the ischemic canine myocardium in vivo is dominated by deoxymyoglobin.

Animals↗

PubMed1999

Integrin alphavbeta3-RGDS interaction mediates fibrin-induced morphological changes of glomerular endothelial cells.

BACKGROUND: In our previous studies, we found that intraglomerular deposition of fibrin and its metabolites was related to glomerular sclerosis and reduced renal function. It has been reported that both overlying and underlying fibrin may induce specific morphological changes of cultured endothelial cells from large blood vessels. The dependency of these morphological changes on the integrin alphavbeta3-arginyl-glycyl-aspartyl-serine (RGDS) interaction is still controversial. We hypothesized that glomerular endothelial cells (GECs) stimulated by fibrin might undergo morphological changes through an integrin alphavbeta3-RGDS interaction. Methods. In vitro studies were performed to examine the growing status of GECs stimulated by overlying and underlying fibrin gels in the presence or absence of the following: 50 microg/ml anti-alphavbeta3 integrin monoclonal antibody 23C6 or nonimmune mouse IgG, 1 mg/ml synthetic RGDS or arginyl-glycyl-glycyl-serine (RGGS) peptide, 10 mg/ml sodium heparin, 100 microg/ml cycloheximide, and 10 microM actinomycin D. Fast protein liquid chromatography (FPLC)-purified fibrinogen and the third to fifth passages of human GECs were also used in this study. RESULTS: GECs developed capillary tube structure after 60 hours of culturing on fibrin gels, and GECs cultured on gelatin-coated plates displayed a monolayer of cobblestone-like cells in the presence or absence of 23C6 and synthetic RGDS peptide. Fibrin-induced capillary tube formation was promoted by 23C6 and inhibited by RGDS peptide, cycloheximide, and actinomycin D. Disorganization of the GEC monolayer was induced by overlying fibrin, but was not induced by overlying agarose gels and glass cover slips or culturing in fibrinogen, 0.05 NIH U/ml thrombin, fibrin supernatants, as well as in fibrin degradation products. Disorganization of GEC monolayer can be induced by both des-AA-fibrin and des-AABB-fibrin and was unaffected by heparin. Furthermore, both 23C6 and synthetic RGDS peptide prevented disorganization of GECs induced by overlying fibrin, whereas nonimmune mouse IgG, synthetic RGGS peptide, cycloheximide, and actinomycin D had no similar effect. CONCLUSIONS: GECs cultured on fibrin gels may develop capillary structure spontaneously, and GECs covered by fibrin gels may undergo disorganization. Our data suggest that these GEC morphological changes are mediated by an integrin alphavbeta3-RGDS interaction.

Antibodies, Monoclonal↗

PubMed1999

The transfer of resources from middle-aged children to functionally limited elderly parents: providing time, giving money, sharing space.

The transfer of resources from middle-aged children to their functionally limited elderly parents is addressed from the perspective of the children who may allocate time to caregiving, share household space, and give money to parents. A simultaneous-equations model estimates the extent to which the three modes of transfer are interdependent, given the parents' needs for resources and the children's ability to provide them. Caregiving is the primary mode of resource transfer and is of overriding importance for individuals who depend daily on help from other persons. Coresidence and financial assistance complement direct human assistance and, at the margin, have a substantially large effect on caregiving time. It is, therefore, relevant to consider all modes of transfer in order to better understand how families accommodate the needs of their frail and disabled members.

Aged↗

PubMed1999

Altered drug interaction and regulation of topoisomerase IIbeta: potential mechanisms governing sensitivity of HL-60 cells to amsacrine and etoposide.

Topoisomerase II (topo II), an enzyme essential for cell viability, is present in mammalian cells as the alpha- and beta-isoforms. In human leukemia HL-60/S or HL-60/doxorubicin (DOX)0.05 cells, the levels of topo IIalpha- or beta-protein were similar in either asynchronous exponential or synchronized cultures. Although topo IIalpha was hypophosphorylated in HL-60/DOX0.05 compared with HL-60/S cells, both overall and site-specific hyperphosphorylation of topo IIbeta was apparent in HL-60/DOX0.05 compared with HL-60/S cells. The phosphorylation of topo IIalpha and not beta was enhanced in the S and G(2) + M phases of HL-60/S cells. In contrast, an increase in the phosphorylation of topo IIbeta compared with alpha was apparent in the G(1) and S phases of HL-60/DOX0.05 cells. The cytotoxicity and depletion of topo IIalpha or beta in cells treated with drug for 1 h revealed that mole-for-mole, amsacrine was 2-fold more effective than etoposide in killing HL-60/S or HL-60/DOX0.05 cells and in depleting the beta versus alpha topo II protein. Present results demonstrate that: 1) hyperphosphorylation of topo IIbeta in HL-60/DOX0.05 cells may be a compensatory consequence of the hypophosphorylation of topo IIalpha to maintain normal topo II function during proliferation, and 2) enhanced sensitivity of HL-60/S or HL-60/DOX0.05 cells to amsacrine may be due to the preferential interaction and depletion of topo IIbeta.

Amsacrine↗

PubMed1999

Formation of adeno-associated virus circular genomes is differentially regulated by adenovirus E4 ORF6 and E2a gene expression.

A central feature of the adeno-associated virus (AAV) latent life cycle is persistence in the form of both integrated and episomal genomes. However, the molecular processes associated with episomal long-term persistence of AAV genomes are only poorly understood. To investigate these mechanisms, we have utilized a recombinant AAV (rAAV) shuttle vector to identify circular AAV intermediates from transduced HeLa cells and primary fibroblasts. The unique structural features exhibited by these transduction intermediates included circularized monomer and dimer virus genomes in a head-to-tail array, with associated specific base pair alterations in the 5' viral D sequence. In HeLa cells, the abundance and stability of AAV circular intermediates were augmented by adenovirus expressing the E2a gene product. In the absence of E2a, adenovirus expressing the E4 open reading frame 6 gene product decreased the abundance of AAV circular intermediates, favoring instead the linear replication form monomer (Rfm) and dimer (Rfd) structures. In summary, the formation of AAV circular intermediates appears to represent a new pathway for AAV genome conversion, which is consistent with the head-to-tail concatemerization associated with latent-phase persistence of rAAV. A better understanding of this pathway may increase the utility of rAAV vectors for gene therapy.

Adenovirus E2 Proteins↗

PubMed1999

Functional interactions between herpesvirus oncoprotein MEQ and cell cycle regulator CDK2.

Marek's disease virus, an avian alphaherpesvirus, has been used as an excellent model to study herpesvirus oncogenesis. One of its potential oncogenes, MEQ, has been demonstrated to transform a rodent fibroblast cell line, Rat-2, in vitro by inducing morphological transformation and anchorage- and serum-independent growth and by protecting cells from apoptosis induced by tumor necrosis factor alpha, C2-ceramide, UV irradiation, or serum deprivation. In this report, we show that there is a cell cycle-dependent colocalization of MEQ protein and cyclin-dependent kinase 2 (CDK2) in coiled bodies and the nucleolar periphery during the G1/S boundary and early S phase. To our knowledge, this is the first demonstration that CDK2 is found to localize to coiled bodies. Such an in vivo association and possibly subsequent phosphorylation may result in the cytoplasmic translocation of MEQ protein. Indeed, MEQ is expressed in both the nucleus and the cytoplasm during the G1/S boundary and early S phase. In addition, we were able to show in vitro phosphorylation of MEQ by CDKs. We have mapped the CDK phosphorylation site of MEQ to be serine 42, a residue in the proximity of the bZIP domain. An indirect-immunofluorescence study of the MEQ S42D mutant, in which the CDK phosphorylation site was mutated to a charged residue, reveals more prominent cytoplasmic localization. This lends further support to the notion that the translocation of MEQ is regulated by phosphorylation. Furthermore, phosphorylation of MEQ by CDKs drastically reduces the DNA binding activity of MEQ, which may in part account for the lack of retention of MEQ oncoprotein in the nucleus. Interestingly, the localization of CDK2 in coiled bodies and the nucleolar periphery is observed only in MEQ-transformed Rat-2 cells, implicating MEQ in modifying the subcellular localization of CDK2. Taken together, our data suggest that there is a novel reciprocal modulation between the herpesvirus oncoprotein MEQ and CDK2.

Animals↗