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Biomedical subjects

C C Chang

Publications and source records attributed to C C Chang.

At least 181 records · Page 10Linked to original sources

Genetic characterization of the mRNAs encoding alpha-bungarotoxin: isoforms and RNA editing in Bungarus multicinctus gland cells.

The mRNA encoding alpha-bungarotoxin (alpha-Butx) was prepared from the venom glands of Bungarus multicinctus by Cap-finder cDNA synthesis. The sequences of the 3'- and 5'-flanking regions including a signal peptide of alpha-Butx were almost identical with those of Elapidae and Hydrophiidae toxins, suggesting that they may have the same origin. Sixteen polymorphic mRNA sequences of alpha-Butx were detected from B.multicinctus gland cells. Analysis of the genomic DNA of alpha-Butx indicated that the polymorphic mRNA originated from one DNA sequence. Most of the mutations in alpha-Butx mRNA were silent and the hot-spot variations occurred at 78, 107, 129, 198 and 201 nt in alpha-Butx mRNA. Ten distinct protein sequences of alpha-Butx could be deduced from the polymorphic mRNA and one of the isoforms has already been isolated. Since alpha-Butx DNA is a single copy in the genome, the RNA polymorphism might result from post-transcriptional editing. These results indicate that the authentic alpha-Butx is in fact derived from edited mRNAs. RNA editing may contribute a common mechanism toward the diversity of alpha-neurotoxins in snake glands.

Amino Acid Sequence↗

Homologous protection but lack of heterologous-protection by various species and types of Bartonella in specific pathogen-free cats.

Cat-scratch disease (CSD) is caused by Bartonella henselae, and possibly by B. clarridgeiae. In immuno-compromised persons, B. henselae is one of the agents causing bacillary angiomatosis. Domestic cats are the main reservoir of these bacteria, which are transmitted primarily from cat to cat by fleas. Possible strategies to prevent the spread of infection among cats are to eliminate flea infestation or to prophylactically immunize cats. In order to develop an appropriate vaccine, it is important to determine if cats become resistant to re-infection by the same strain or various types or species of Bartonella. In a series of experiments, 21 SPF cats were experimentally infected by the intradermal route with 10(5)-10(10) colony-forming units/ml of either B. henselae type II (17 cats), or a new strain 'Humboldt' isolated from a mountain lion (4 cats). The cats were bled weekly to every other week for determination of bacteremia and specific antibody production. After they cleared their infection, they were challenged by a homologous or heterologous strain of Bartonella: 10 cats were challenged with B. henselae type II, three cats with B. henselae type I, four cats with B. clarridgeiae and four cats with the 'Humboldt' strain. Seven of these cats received a third inoculum dose resulting in three cats sequentially infected with sequence B. henselae type II/B. henselae type II/'Humboldt', two cats with sequence B. henselae type II/'Humboldt'/B. clarridgeiae, and two cats with the sequence 'Humboldt'/B. henselae type II/'Humboldt'. All cats challenged with a homologous strain remained abacteremic after challenge and had an increased IgG antibody titer. All cats challenged with either a different Bartonella species or type became bacteremic. The few cats receiving a third inoculum with a strain homologous to the initial strain remained abacteremicafter that challenge. All cats infected with B. clarridgeiae suffered relapsing bacteremia compared to only 36% of the B. henselae infected cats and 22% of the 'Humboldt'-infected cats (p=0.008). The duration of bacteremia was significantly longer in B. henselae primary-infected cats (mean: 34 weeks) than B. henselae heterologously challenged cats (mean: 9 weeks) (p=0.014). These data clearly indicate the lack of cross-protection between B. henselae and B. clarridgeiae and furthermore, indicate the lack of protection between B. henselae types I and II, and a wildlife isolate. A vaccine strategy for CSD prevention in domestic cats will require a multivalent vaccine approach.

Animals↗

Cloning and functional expression of B chains of beta-bungarotoxins from Bungarus multicinctus (Taiwan banded krait).

The cDNA species encoding the B chains (B1 and B2) of beta-bungarotoxins (beta-Bgt) were constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus (Taiwan banded krait). The deduced amino acid sequences of the B chains were different from those determined previously by a protein sequencing technique. One additional Arg residue is inserted between Val-19 and Arg-20 of the B1 chain. Similarly the insertion of one additional Val residue between Val-19 and Arg-20 of the B2 chain is noted. Thus the B chains should comprise 61 amino acid residues. Moreover, the residues at positions 44-46 are Gly-Asn-His, in contrast with a previous result showing the sequence His-Gly-Asn. Instead of Asp, the residues at positions 41 and 43 are Asn. The B chain was subcloned into the expression vector pET-32a(+) and transformed into Escherichia coli strain BL21(DE3). The recombinant B chain was expressed as a fusion protein and purified on a His-Bind resin column. The yield of affinity-purified fusion protein was increased markedly by replacing Cys-55 of the B chain with Ser. However, the isolated B(C55S) chain became insoluble in aqueous solution after removal of the fused protein from the affinity-purified product, suggesting that protein-protein interactions might be crucial for stabilizing the structure of the B chain. The B(C55S) chain fusion protein showed activity in blocking the voltage-dependent K+ channel, but did not inhibit the binding of beta-Bgt to synaptosomal membranes. These results, together with the finding that modification of His-48 of the A chain of beta-Bgt caused a marked decrease in the ability to bind toxin to its acceptor proteins, suggest that the B chain is involved in the K+ channel blocking action observed with beta-Bgt, and that the binding of beta-Bgt to neuronal receptors is not heavily dependent on the B chain.

Amino Acid Sequence↗

Improvement of heterologous protein productivity using recombinant Yarrowia lipolytica and cyclic fed-batch process strategy.

A cyclic fed-batch bioprocess is designed and a significant improvement of rice alpha-amylase productivity of recombinant Yarrowia lipolytica is illustrated. A bioprocess control strategy developed and reported here entails use of a genetically stable recombinant cloned for heterologous protein, use of optimized media for cell growth and enzyme production phases, and process control strategy enabling high cell-density culture and high alpha-amylase productivity. This process control can be achieved through maintaining a constant optimal specific cell growth rate at a predetermined value (i.e., 0.1 h-1), controlling medium feed rate commensurate with the cell growth rate, and maintaining a high cell-density culture (i.e., 60-70 g/L) for high productivity of cloned heterologous protein. The volumetric enzyme productivity (1, 960 units/L. h) achieved from the cyclic fed-batch process was about 3-fold higher than that of the fed-batch culture process (630 units/L. h).

Bioreactors↗

Cooperative effects of v-myc and c-Ha-ras oncogenes on gap junctional intercellular communication and tumorigenicity in rat liver epithelial cells.

The objective of this study was to isolate and partially characterize several rat liver epithelial cell clones containing myc, ras and myc/ras oncogenes in order to study their roles in apoptosis and to test the hypothesis that gap junctional intercellular communication is necessary for apoptosis in solid tissues and that the loss of junctional communication leads to tumorigenesis. The co-transfection of the myc and ras oncogenes in the normal rat liver epithelial cell line (WB-F344) resulted in a loss of functional channels and normal growth regulation; cell-cell communication was significantly decreased and tumorigenicity determined in adult male F344 rats was induced. We examined cell growth properties, gap junctional intercellular communication (GJIC), using the scrape-loading-dye transfer and fluorescence-redistribution-after-photobleaching assays, and tumorigenicity in a series of normal and v-myc-, c-Ha-ras- and v-myc/c-Ha-ras-transfected WB-F344 cell lines. The c-Ha-ras- and the v-myc/c-Ha-ras-transduced cell lines appeared distinctly different from the other lines, having spindle-shaped morphology, shorter generation time and contact insensitivity. On the other hand, the normal WB-F344 cell line and the v-myc-transduced cell line showed excellent GJIC. Moreover, the c-Ha-ras-transduced cell lines displayed decreasing levels of GJIC associated with their increasing tumorigenicity. The v-myc/c-Ha-ras-transformed cell lines showed the lowest levels of GJIC and were also the most tumorigenic. These findings suggest that the reduction of GJIC in c-Ha-ras- and v-myc/c-Ha-ras-transformed WB-F344 cells is linked to their tumorigenic potential. These cell lines should provide valuable tools to study the role of GJIC in apoptosis during tumorigenesis.

Animals↗

Unfolding/folding studies on cobrotoxin from Taiwan cobra venom: pH and GSH/GSSG govern disulfide isomerization at the C-terminus.

Refolding of cobrotoxin was assessed by the exposure degree of its single Trp determined by an acrylamide quenching study. The change in the accessibility of Trp for acrylamide quantitatively reflected the formation of folded cobrotoxin, and the data were confirmed by HPLC and gel electrophoresis analyses. However, the site-specific information provided by quenching Trp fluorescence revealed that the ordered structure in the neighborhood of Trp was attained prior to the complete formation of the tertiary structure of cobrotoxin. HPLC analyses showed that, in addition to refolded cobrotoxin, two novel species (cobrotoxin II and cobrotoxin III) with isomerization of disulfide bonds at the C-terminus of the toxin molecule were produced along the folding reaction. The disulfide pairings in cobrotoxin II and cobrotoxin III were Cys43-Cys55 and Cys54-Cys60 and Cys43-Cys60 and Cys54-Cys55, respectively. Among the three possible two-disulfide species at the C-terminus, the disulfide linkages Cys43-Cys60 and Cys54-Cys55 of cobrotoxin III caused a marked decrease in lethality and resulted in a conformation which was notably different from that observed with the native toxin molecule as evidenced by CD spectra. The refolding reaction was accelerated by the addition of GSH/GSSG, and the resulting products were mostly folded cobrotoxin. However, if GSH/GSSG was not added into the initial folding materials, the yields of cobrotoxin II and cobrotoxin III greatly increased. The conversion of cobrotoxin to its isomers was to be irreversible and pH-dependent: the higher the pH, the faster the rate of conversion. However, this conversion could be partly inhibited by GSH/GSSG. Cobrotoxin II and cobrotoxin III were purified from Taiwan cobra venom as well, and their yields in comparison to that of cobrotoxin in venom were similar to that noted with the folded products in the presence of GSH/GSSG. Moreover, the rate of disulfide isomerization was expected to be slow in venom fluid in which the pH was approximately pH 6.2. Thus, the finding that cobrotoxin represents the predominant neurotoxin species in Taiwan cobra venom is probably associated with the synergistic effects of GSH/GSSG and pH.

Amino Acid Sequence↗

Isoorientin-6"-O-glucoside, a water-soluble antioxidant isolated from Gentiana arisanensis.

The antioxidant activities of isoorientin-6"-O-glucoside were studied using various models. Isoorientin-6"-O-glucoside was more potent than Trolox, probucol and butylated hydroxytoluene (BHT) in reducing the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH). It also scavenged superoxide anion, peroxyl and hydroxyl radicals that were generated by xanthine/xanthine oxidase, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) and Fe3+-ascorbate-EDTA-H2O2 system, respectively. The IC50 value, stoichiometry factor and second-order rate constant were 9.0+/-0.8 microM, 1.8+/-0.1 and 2.6 X 10(10) M(-1) s(-1) for superoxide generation, peroxyl and hydroxyl radicals. However, isoorientin-6"-O-glucoside did not inhibit xanthine oxidase activity or scavenge hydrogen peroxide (H2O2), carbon radical or 2,2'-azobis(2,4-dimethyl-valeronitrile) (AMVN)-derived peroxyl radical in hexane. Isoorientin-6"-O-glucoside inhibited Cu2+-induced oxidation of human low-density lipoprotein (LDL) as measured by fluorescence intensity, thiobarbituric acid-reactive substance formation and electrophoretic mobility. Since isoorientin-6"-O-glucoside did not possess pro-oxidant activity, it may be an effective water-soluble antioxidant that can prevent LDL against oxidation.

Antioxidants↗

Molecular cloning of YlPMR1, a S. cerevisiae PMR1 homologue encoding a novel P-type secretory pathway Ca2+ -ATPase, in the yeast Yarrowia lipolytica.

A novel P-type ATPase gene, Saccharomyces cerevisiae PMR1 homologue (YlPMR1), has been cloned and sequenced in the yeast, Yarrowia lipolytica. The putative gene product has 928 amino acids with a calculated molecular mass of 100050 Da and a pI of 5.15. The deduced amino-acid sequence analysis demonstrated that the cloned gene product contains all 10 of the conserved regions in P-type ATPases and exhibits 55% amino-acid identity to the S. cerevisiae PMR1 gene product; however, it shows a relatively lower homology to PMCA (24%) and SERCA (33%), confirming the presence of a third class of Ca2+-ATPase (secretory pathway Ca2+-ATPase, SPCA). The YlPMR1-disrupted strain shows defective growth in low Ca2+ or EGTA-containing medium. In fact, a longer lag time (60 h) was observed in YlPMR1-defective mutant cells during cultivation in EGTA-containing YPD medium. These growth defects were overcome by adding Ca2+ and Mn2+ into the medium. Interestingly, whereas Mn2+ inhibits growth of the control strain, it significantly improves the growth of YlPMR1-disrupted cells. These results suggest an involvement of the YlPMR1 gene product in Ca2+ and Mn2+ ion homeostasis in Y. lipolytica.

Amino Acid Sequence↗

Neoplastic phenotype of gap-junctional intercellular communication-deficient WB rat liver epithelial cells and its reversal by forced expression of connexin 32.

Gap-junctional intercellular communication (GJIC) is involved in cellular growth control and is often reduced in neoplastic cells. In this study, four GJIC-deficient rat liver epithelial cell lines (WB-aB1, WB-bA2, WB-cD6, and WB-dA2) were examined for altered growth and tumorigenicity in comparison with their GJIC-competent parental cell line, WB-F344. WB-aB1 cells were also forced to express connexin 32 (Cx32) by transduction with a Cx32 cDNA retroviral expression vector to help determine whether the restoration of GJIC could reverse their neoplastic phenotype. WB-aB1 and WB-bA2 cells had faster population doubling times (PDTs) and higher saturation densities (SDs) than did WB-F344 cells. In contrast, the growth of WB-cD6 and WB-dA2 cells was not significantly different from that of WB-F344 cells. WB-aB1 and WB-bA2 cells formed tumors in male F344 rats, but WB-cD6 and WB-dA2 cells did not. After transduction of WB-aB1 cells with Cx32, four stable clones (WB-a/32-3, -8, -9, and -10) were isolated that had GJIC levels of 5.2%, 44.5%, 69.8%, and 90.5%, respectively. The growth of poorly coupled clones 3 and 8 was similar to that of parental WB-aB1 cells, but the growth of well-coupled clones 9 and 10 was similar to that of WB-F344 cells. The tumorigenicity of WB-a/32-9 and WB-a/32-10 cells was also significantly lower than that of WB-aB1 cells. Our results suggest that reduced GJIC contributes to neoplastic transformation of WB cells, that additional changes are necessary, and that restoration of GJIC by forced Cx32 protein expression can suppress the neoplastic phenotype of these cells.

Animals↗

Positron emission tomography for detecting iodine-131 nonvisualized metastasis of well-differentiated thyroid carcinoma: two case reports.

To investigate the usefulness of positron emission tomography (PET) in detecting I-131 nonvisualized metastatic foci of well-differentiated thyroid carcinoma (WDTC), 2 patients with papillary and follicular thyroid carcinoma respectively, were studied with I-131 total body scan, thallium-201 scan, Tc99m bone scan and [18F]-2-deoxy-2-fluoro-D-glucose (FDG) PET. Case 1 showed no metastatic lesion in I-131 (up to 150 mCi) total body scan, 1 anterior mass in thallium-201 scan, none in Tc99m bone scan and 7 including the main anterior mediastinal mass in FDG-PET. Case 2 showed 2 metastatic lesions in I-131 (150 mCi) total body scan, 11 bony metastatic lesions in Tc99m bone scan and 13 in FDG-PET. However, lower extremities were not scanned in FDG-PET. Tumor/background ratio of 1.5 or above is needed to be visualized grossly. The FDG-PET tumor/background ratios are higher than those of thallium-201 except in one site. In conclusion, I-131 scintigraphy is still the first line method to use in detecting WDTC recurrence and metastasis, as I-131 has the advantage of being both a therapeutic and imaging agent. For I-131 nonvisualized metastasis of WDTC, thallium scintigraphy and FDG-PET may be considered. Even though FDG-PET has better sensitivity, resolution imaging and spatial localization, this has to be balanced with its higher cost when compared with thallium scintigraphy.

Adenocarcinoma, Follicular↗

Recombinant bioprocess optimization for heterologous protein production using two-stage, cyclic fed-batch culture.

A two-stage, cyclic fed-batch bioprocess was designed, and its performance evaluated to improve rice alpha-amylase productivity by the yeast Yarrowia lipolytica SMY2 (MatA, ade1, ura3, xpr2), ATCC 201847, containing a replicative plasmid coding for a rice alpha-amlyase. Transcription of the recombinant gene is controlled by the XPR2 promoter. The first stage (or growth stage) was operated in the fed-batch mode, and the growth medium, designed to maintain a constant high cell density (i.e., 60 g/l), was fed according to a predetermined and preprogrammed optimal feed rate which, in turn, maintained the specific cell growth rate at an optimal value (i.e., 0.1 h-1). Typically, when the volume in the first stage reached a preset value, a portion of culture broth (i.e., 55%) was transferred to the second stage (or production stage). The remaining cells in the growth stage were then fed with fresh growth medium according to the bioprocess control strategy developed, while induction of alpha-amylase expression and its production was taking place in the second stage. The second stage was also operated in the fed-batch mode, and the production medium designed to maintain a constant high cell density and high productivity of heterologous protein was fed at a predetermined and preprogrammed rate, which maintained the specific cell growth rate at an optimal level. The volumetric alpha-amylase productivity achieved (1835 units l-1 h-1) from the two-stage, cyclic fed-batch culture process was twofold higher than that of the fed-batch culture process. The genetic stability of the recombinant strain and the design of optimal media for growth and production stages are also critically important to a successful implementation of the two-stage, cyclic fed-batch process for production of heterologous protein.

Recombinant Proteins↗

Cerebral blood flow measurement in patients with impaired consciousness: usefulness of 99mTc-HMPAO single-photon emission tomography in clinical practice.

The relationship between impairment of consciousness and quantitative cerebral blood flow (CBF) was investigated. The mean CBF of the whole brain was measured by the Patlak-plot method using technetium-99m hexamethylpropylene amine oxime single-photon emission tomography (99mTc-HMPAO SPET) in patients with the following diseases: cerebral infarction, intraparenchymal haemorrhage, subarachnoid haemorrhage, brain tumour and cerebral contusion. The clinical symptoms were evaluated according to the severity of impaired consciousness, aphasia and dementia. Four hundred and eighty-five CBF measurements were performed. Patients with alert consciousness showed an age-related decline in mean CBF. Patients with aphasia showed a significant reduction in mean CBF compared with those without aphasia. Impaired consciousness was proportional to reduction in mean CBF regardless of types of pathology, and the size of lesion did not influence the mean CBF. Patients with dementia showed a significant reduction in mean CBF proportional to the severity of dementia. The quantitative measurement of CBF using 99mTc-HMPAO SPET is reliable in clinical evaluations.

Adolescent↗

Primary hyperparathyroidism in pregnancy--report of 3 cases.

Three cases of primary hyperparathyroidism in pregnancy are described. Patient 1 developed left thigh pain and lower abdominal pain at 34 weeks' gestation. Patient 2 had right flank pain and lower abdominal pain at 32 weeks' gestation. Both patients accepted medical therapy initially, which resulted in poor control of hypercalcemia. Patient 1 delayed her parathyroidectomy until the postpartum period; she had maternal hypercalcemia and neonatal hypocalcemia. Patient 2 accepted parathyroidectomy at 32 weeks' gestation with an uneventful outcome for both mother and baby. Patient 3 was asymptomatic; her hyperparathyroidism was diagnosed postpartum after neonatal hypocalcemia and agreed to parathyroidectomy. All 3 patients had a parathyroid adenoma.

Adenoma↗

Novel inhibition of contractility by wortmannin in skeletal muscle.

1. The effects of wortmannin and 2-(4-morpholinyl)-8-phenyl-1[4H]-benzopyran-4-one (LY294002), inhibitors of phosphatidylinositol 3-kinase, on the contractile responses of murine skeletal muscle were studied. Wortmannin (10-100 microM) suppressed twitch and tetanic contraction evoked by field stimulation of diaphragm without causing elevation of muscle tone. The inhibition was quasi-irreversible with IC50 approximately 15 microM. In contrast, LY294002 increased twitch responses and elevated muscle tone. 2. Wortmannin reversibly depressed the maximal slope of action potential upstroke by approximately 40% and inhibited the membrane depolarization and spontaneous burst of action potential induced by crotamine, a polypeptide toxin that activates the Na+ channel of skeletal muscle. 3. Wortmannin inhibited contractures evoked by high K+, ryanodine and caffeine, but potentiated the contracture induced by rapamycin, which binds to myoplasmic FK506 binding protein, an immunophilin closely associated with the ryanodine receptor. The contractures elicited by cardiotoxin, which disrupts the integrity of sarcolemma and thereby elevates 'myoplasmic' Ca2+ level, were suppressed only slightly. 4. In placed left atrium and ventricular strip, wortmannin and LY294002 produced a positive inotropic effect. 5. The results suggest that, in addition to depressing the Ca2+ mobilization from sarcoplasmic reticulum, wortmannin exerts a novel inhibitory action on the excitation-contraction coupling in skeletal muscle but not in cardiac muscle.

Action Potentials↗

Phenotype-genotype relationships in complementation group 3 of the peroxisome-biogenesis disorders.

The peroxisome-biogenesis disorders (PBDs) are a set of often lethal genetic diseases characterized by mental retardation and defective peroxisomal matrix protein import. Mutations in PEX12 are known to underlie the disease in two patients from complementation group 3 of the PBDs. Here we show that all patients from this group carry mutations on both alleles of PEX12. A comparison between PEX12 genotypes and the clinical and cellular phenotypes of the corresponding PBD patients suggests a relatively straightforward relationship between genotype and phenotype in this group of the PBDs, such that the loss of PEX12 function leads to more-severe cellular and clinical phenotypes. However, one patient who presented relatively mild clinical and cellular phenotypes was a compound heterozygote for two seemingly severe mutations on each PEX12 allele. PEX12 mRNA present in the patient's cells was derived from only one allele, the one that carried a 2-bp deletion early in the PEX12 coding region, c.26,27Delta. The deduced protein product of this mRNA would contain only the first eight amino acids of the protein, and yet this mutant PEX12 cDNA displayed significant PEX12 activity in a functional complementation assay. Surprisingly, the PEX12/c.26, 27Delta cDNA directed the synthesis of a 29-kD PEX12 protein in vitro, a result that is consistent with translation initiation at a downstream AUG codon. Transfection studies confirmed the expression of similarly sized PEX12 proteins from the PEX12/c.26,27Delta allele. Thus, it appears that translation initiation at internal AUG codons may modulate disease phenotypes and should be considered whenever unexpectedly mild phenotypes result from severe mutations early in the coding region.

Amino Acid Sequence↗

Deregulation of cdc2 gene expression correlates with overexpression of a 110 kDa CCAAT box binding factor in transformed cells.

Eukaryotic cell cycle progression is regulated by an orderly and sequential activation of several cyclin-dependent kinases, which phosphorylate key substrates during this process. p34cdc2, the catalytic subunit of cdc2 kinase, is expressed at the late G1/S boundary and is required for the G2-->M phase transition. Transactivation of the human cdc2 promoter by the DNA tumor virus-encoded oncogenic protein SV40 large T antigen is mediated by induction of a novel 110 kDa CCAAT box binding factor (CBF/cdc2). To investigate whether induction of CBF/cdc2 is an intrinsic property of the viral oncoprotein or is a common event during transformation of normal cells, expression of CBF/cdc2 was analyzed in many human tumor cell lines and in rodent cells spontaneously transformed or stably expressing various oncogenes. Our results showed that CBF/cdc2 was overexpressed in all transformed cells examined, including human 293, MCF-7, HeLa and HepG2 cells. Moreover, expression of CBF/cdc2 was elevated in spontaneously transformed rat liver epithelial cells (C4T), but not detectable in the non-tumorigenic parental (RLE) cells. The elevated levels of CBF/cdc2 expression in C4T cells correlated well with increased cdc2 mRNA and p34cdc2 levels. CBF/cdc2 was also overexpressed in a rat liver epithelial cell line (WB) stably transfected with various oncogenes, v-myc, v-Ha-ras and mutated rat neu and v-src. Using an electrophoretic mobility shift assay, specific binding of CBF/cdc2 to the CCAAT box motifs of the human cdc2, cycA and cdc25C promoters was detected, suggesting that transcription of these cell cycle regulatory genes are coordinately activated by CBF/cdc2.

Animals↗