Biomedical subjects

T Toda

Publications and source records attributed to T Toda.

At least 73 records · Page 4Linked to original sources

PubMed2000

Rho-dependence of Schizosaccharomyces pombe Pck2.

BACKGROUND: In metazoans, the HR1 domain, a motif found in a number of proteins including the protein kinase C-related PRKs, is responsible for an interaction with Rho-GTPases. The structural similarity between the Schizosaccaromyces pombe Pck proteins and the mammalian Rho-dependent protein kinase C-related family, has led us to investigate the relationship between the function of Rho and that of Pck1/2. RESULTS: Rho1 is shown to interact with the conserved N-terminal HR1 domain of Pck1/2 in vitro and in vivo. Lethal overproduction of Rho1 is neutralized by co-expression of the Pck2 HR1 domain, which by itself compromises growth when overproduced. The Pck2-Rho1 interaction has a profound effect on the steady state expression of Pck2 and this is shown to parallel the immunoprecipitated activity and phosphorylation of Pck2 at its activation loop site. It is further shown that Pck2 becomes localized at the septum, where Rho1 is also located. CONCLUSIONS: The results demonstrate that the Pck proteins are Rho1 effectors in fission yeast and that the HR1 domain is a universal motif for the Rho-GTPase interaction. Furthermore, the evidence supports the contention that the yeast Pck1 and Pck2 proteins are primitive protein kinases, which in vertebrates have evolved into the two distinct PKC and PRK families.

Amino Acid Sequence↗

PubMed2000

A comparison of the relative efficacies of four hand and rotary instrumentation techniques during endodontic retreatment.

AIM: The purpose of this study was to quantify the amount of remaining gutta-percha/scaler on the walls of root canals when two engine-driven instruments (Quantec and ProFile) and two hand instruments (K-file and Hedström file) were used to remove these materials. The amount of apically extruded debris and the time required for treatment were also recorded. METHODOLOGY: One hundred extracted mandibular premolars were prepared using a modified step-back, flare technique and obturated with the lateral condensation technique. After repreparation with the test instruments, the specimens were cut transversally at the cervical, middle and apical thirds with steel discs and the three sections were split longitudinally. The amount of residual debris on the canal walls in each section was examined using a stereomicroscope. RESULTS: In all groups the cervical and middle thirds showed no debris. In the apical third, obturating material was observed in some specimens. No statistically significant difference was found between the two groups for incidence of debris, although the Hedström group showed a greater number of samples with remaining gutta-percha/sealer. When analysing dirty specimens only, there was a statistically significant difference between the four groups (P < 0.01) with the Hedström group having significantly less length of canal wall with remaining obturation material than the Quantec group. There was no significant difference amongst the groups for weight of extruded debris. However, there was a significant difference amongst the groups for mean treatment time with the Hedström file group requiring significantly less time than the Quantec group (P < 0.001); no significant differences were found between the other groups. Six instruments fractured in the Quantec group, four in the ProFile group, two in the Hedström group and two in the K-type group. CONCLUSIONS: The results showed that overall, all instruments may leave filling material inside the root canal. During retreatment there is a risk of instrument breakage, especially rotary instruments.

Analysis of Variance↗

PubMed2000

Expression of p53, bcl-2, and bax as predictors of response to radiotherapy in esophageal cancer.

The sensitivity of cancers to radiotherapy or chemotherapy may be influenced by susceptibility to apoptosis. We evaluated whether expression of three proteins regulating apoptosis, p53, bcl-2, and bax, could predict the effect of radiotherapy in esophageal cancers. We used immunohistochemical staining for these protein regulators of apoptosis to study biopsy specimens obtained from 25 patients with esophageal squamous cell carcinoma before they underwent preoperative radiotherapy. Effectiveness of radiotherapy was assessed by barium esophagography, esophagoscopy, and computed tomography. Radiotherapy was effective in 12 patients and ineffective in 13 patients. Biopsy specimens from the 25 patients showed expression of p53, bcl-2, and bax to be 48.0%, 32.0%, and 76.0% respectively. Effectiveness of radiotherapy was correlated with p53 expression (p = 0.047), but bcl-2 and bax expression showed no relationship to effectiveness of radiotherapy. Expression of p53 protein in biopsy specimens may predict effectiveness of preoperative radiotherapy in esophageal cancers.

Aged↗

PubMed2000

Schizosaccharomyces pombe rho2p GTPase regulates cell wall alpha-glucan biosynthesis through the protein kinase pck2p.

Schizosaccharomyces pombe rho1(+) and rho2(+) genes are involved in the control of cell morphogenesis, cell integrity, and polarization of the actin cytoskeleton. Although both GTPases interact with each of the two S. pombe protein kinase C homologues, Pck1p and Pck2p, their functions are distinct from each other. It is known that Rho1p regulates (1,3)beta-D-glucan synthesis both directly and through Pck2p. In this paper, we have investigated Rho2p signaling and show that pck2 delta and rho2 delta strains display similar defects with regard to cell wall integrity, indicating that they might be in the same signaling pathway. We also show that Rho2 GTPase regulates the synthesis of alpha-D-glucan, the other main structural polymer of the S. pombe cell wall, primarily through Pck2p. Although overexpression of rho2(+) in wild-type or pck1 delta cells is lethal and causes morphological alterations, actin depolarization, and an increase in alpha-D-glucan biosynthesis, all of these effects are suppressed in a pck2 delta strain. In addition, genetic interactions suggest that Rho2p and Pck2p are important for the regulation of Mok1p, the major (1-3)alpha-D-glucan synthase. Thus, a rho2 delta mutation, like pck2 delta, is synthetically lethal with mok1-664, and the mutant partially fails to localize Mok1p to the growing areas. Moreover, overexpression of mok1(+) in rho2 delta cells causes a lethal phenotype that is completely different from that of mok1(+) overexpression in wild-type cells, and the increase in alpha-glucan is considerably lower. Taken together, all of these results indicate the presence of a signaling pathway regulating alpha-glucan biosynthesis in which the Rho2p GTPase activates Pck2p, and this kinase in turn controls Mok1p.

Cell Wall↗

PubMed2000

Isoflavone-rich soy protein isolate attenuates bone loss in the lumbar spine of perimenopausal women.

BACKGROUND: No published studies have directly examined the effect of soy protein with isoflavones on bone or bone turnover in perimenopausal women. OBJECTIVE: Our objective was to determine the effects of 24 wk of consumption of soy protein isolate with isoflavones (80.4 mg/d) in attenuating bone loss during the menopausal transition. DESIGN: Perimenopausal subjects were randomly assigned, double blind, to treatment: isoflavone-rich soy (SPI+; n = 24), isoflavone-poor soy (SPI-; n = 24), or whey (control; n = 21) protein. At baseline and posttreatment, lumbar spine bone mineral density (BMD) and bone mineral content (BMC) were measured by using dual-energy X-ray absorptiometry. At baseline, midtreatment, and posttreatment, urinary N:-telopeptides and serum bone-specific alkaline phosphatase (BAP) were measured. RESULTS: The percentage change in lumbar spine BMD and BMC, respectively, did not differ from zero in the SPI+ or SPI- groups, but loss occurred in the control group (-1.28%, P: = 0.0041; -1.73%, P: = 0.0037). By regression analysis, SPI+ treatment had a positive effect on change in BMD (5.6%; P: = 0.023) and BMC (10.1%; P: = 0.0032). Baseline BMD and BMC (P: < or = 0.0001) negatively affected the percentage change in their respective models; baseline body weight (P: = 0.0036) and bone-free lean weight (P: = 0.016) contributed positively to percentage change in BMD and BMC, respectively. Serum BAP posttreatment was negatively related to percentage change in BMD (P: = 0.0016) and BMC (P: = 0.019). Contrast coding using analyses of covariance with BMD or BMC as the outcome showed that isoflavones, not soy protein, exerted the effect. CONCLUSION: Soy isoflavones attenuated bone loss from the lumbar spine in perimenopausal women.

Adult↗

PubMed2000

Inhibitory effects of Bifidobacterium-fermented soy milk on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced rat mammary carcinogenesis, with a partial contribution of its component isoflavones.

High consumption of soybean and soybean-related products is hypothesized to contribute to protection against breast cancer. Soybean is a rich source of genistein, a putative cancer chemopreventive agent. Fermented soy milk (FSM), which is made of soy milk fermented with the Bifidobacterium breve strain Yakult, contains larger amounts of the isoflavone aglycones genistein and daidzein than unfermented soy milk. In the present study, we examined the effects of FSM and its component isoflavone mixture (genistein:daidzein 4:1) on 2-amino-1-methyl-6-phenylimidazo[4, 5-b]pyridine (PhIP)-induced mammary carcinogenesis in rats. Starting at 7 weeks of age, female Sprague-Dawley rats were given PhIP at a dose of 85 mg/kg body wt by intragastric administration four times a week for 2 weeks. They were fed control high fat basal diet or experimental high fat diet containing 10% FSM or 0.02 or 0.04% isoflavone mixture during and after carcinogen exposure. The incidences (percentage of rats with tumors) of mammary gland tumors were 71% in the control diet group, 51% in the FSM group and 68 and 61% in the groups treated with isoflavone mixture at 0.02 and 0.04%, respectively. Mammary tumor multiplicities (number of tumors per rat) were 1.2 +/- 0.2 for 10% FSM, 2.2 +/- 0.4 for 0.02% isoflavone mixture and 1.5 +/- 0.3 for 0.04% isoflavone mixture, being clearly smaller than the control diet value (2.6 +/- 0.5). Furthermore, feeding of FSM and the isoflavone mixture at both doses reduced the sizes of mammary tumors. Since the amounts of isoflavones in 10% FSM are approximately equivalent to those in the 0.02% isoflavone mixture, the chemopreventive activity of FSM could be partly attributable to the presence of isoflavones such as genistein and daidzein.

Animals↗

PubMed2000

The Drosophila embargoed gene is required for larval progression and encodes the functional homolog of schizosaccharomyces Crm1.

The CRM1 (Exportin 1) protein is a receptor for leucine-rich nuclear export signal sequences. We have molecularly characterized the Drosophila melanogaster embargoed (emb) gene and find that it encodes a product with 49 and 71% sequence identity to the fission yeast Schizosaccharomyces pombe and human CRM1 proteins, respectively. We show that expression of the emb cDNA is sufficient to suppress the growth phenotype of both conditional-lethal and null S. pombe crm1(-) mutant strains, suggesting that emb encodes the functional homologue of the S. pombe Crm1 protein. Through mutagenesis screens we have recovered a series of recessive lethal emb mutations. There is a substantial maternal contribution of emb mRNA and animals hemizygous for our emb alleles can develop to second instar larvae but persist at this stage and consistently fail to undergo the molt to the third instar stage. We see a nuclear accumulation of endogenous actin in the intestinal epithelial cells of the emb mutant larvae, consistent with a role for the emb gene product in nuclear export of actin protein.

Actins↗

PubMed2000

The cofactor-dependent pathways for alpha- and beta-tubulins in microtubule biogenesis are functionally different in fission yeast.

The biogenesis of microtubules in the cell comprises a series of complex steps, including protein-folding reactions catalyzed by chaperonins. In addition a group of evolutionarily conserved proteins, called cofactors (A to E), is required for the production of assembly-competent alpha-/beta-tubulin heterodimers. Using fission yeast, in which alp11(+), alp1(+), and alp21(+), encoding the homologs for cofactors B, D, and E, respectively, are essential for cell viability, we have undertaken the genetic analysis of alp31(+), the homolog of cofactor A. Gene disruption analysis shows that, unlike the three genes mentioned above, alp31(+) is dispensable for cell growth and division. Nonetheless, detailed analysis of alp31-deleted cells demonstrates that Alp31(A) is required for the maintenance of microtubule structures and, consequently, the proper control of growth polarity. alp31-deleted cells show genetic interactions with mutations in beta-tubulin, but not in alpha-tubulin. Budding yeast cofactor A homolog RBL2 is capable of suppressing the polarity defects of alp31-deleted cells. We conclude that the cofactor-dependent biogenesis of microtubules comprises an essential and a nonessential pathway, both of which are required for microtubule integrity.

Amino Acid Sequence↗

PubMed2000

Suppressive effects of dietary genistin and daidzin on rat prostate carcinogenesis.

High intake of phytoestrogens through soybeans and their products is thought to be associated with low incidences of prostate and / or breast cancer in Asian countries. Possible chemopreventive effects of genistin or daidzin on rat prostate carcinogenesis were therefore investigated. Male F344 rats were given 10 biweekly subcutaneous injections of 3,2'-dimethyl-4-aminobiphenyl (DMAB) and then either genistin or daidzin in the diet at a concentration of 0.1% for 40 weeks. Other groups of rats given DMAB were treated with genistin or daidzin together with a high dose of testosterone propionate (TP). Both genistin and daidzin reduced the numbers of ventral prostate carcinomas (P < 0.05), with a tendency for decrease in incidence. Invasive carcinomas which developed in the anterior prostate and seminal vesicles with TP were, however, not influenced by the two isoflavones. Thus, the present data suggest that genistin and daidzin possess anti-cancer effects at relatively early stages of prostate cancer development, providing experimental support for epidemiological findings.

Aminobiphenyl Compounds↗

PubMed2000

Cid1, a fission yeast protein required for S-M checkpoint control when DNA polymerase delta or epsilon is inactivated.

The S-M checkpoint is an intracellular signaling pathway that ensures that mitosis is not initiated in cells undergoing DNA replication. We identified cid1, a novel fission yeast gene, through its ability when overexpressed to confer specific resistance to a combination of hydroxyurea, which inhibits DNA replication, and caffeine, which overrides the S-M checkpoint. Cid1 overexpression also partially suppressed the hydroxyurea sensitivity characteristic of DNA polymerase delta mutants and mutants defective in the "checkpoint Rad" pathway. Cid1 is a member of a family of putative nucleotidyltransferases including budding yeast Trf4 and Trf5, and mutation of amino acid residues predicted to be essential for this activity resulted in loss of Cid1 function in vivo. Two additional Cid1-like proteins play similar but nonredundant checkpoint-signaling roles in fission yeast. Cells lacking Cid1 were found to be viable but specifically sensitive to the combination of hydroxyurea and caffeine and to be S-M checkpoint defective in the absence of Cds1. Genetic data suggest that Cid1 acts in association with Crb2/Rhp9 and through the checkpoint-signaling kinase Chk1 to inhibit unscheduled mitosis specifically when DNA polymerase delta or epsilon is inhibited.

Amino Acid Sequence↗

PubMed2000

Variability of bio-optical factors influencing the seasonal attenuation of ultraviolet radiation in temperate coastal waters of Japan

The study identifies the relative contribution of various bio-optical factors to the total attenuation of ultraviolet radiation (UVR) wavelengths and photosynthetically active radiation (PAR) in temperate coastal waters of Japan by surveying the physical properties of the water column, UVR and PAR penetration, and the absorption characteristics of dissolved and particulate material. Spectral absorbance properties of pigment (aph), detritus (ad) and chromophoric dissolved organic material (aCDOM) displayed both seasonal and wavelength specific variability. On an annual basis, absorbance by aCDOM was the highest absorbing fraction (47-59%) for the UVR wavelengths measured (305, 320, 340 and 380 nm) but decreased (32%) at 450 nm. Contribution of pigments to total absorbance was highest (40-60%) during a spring bloom for both UVR and PAR. A large variability (C.V. > 42%) for annual average attenuation coefficients (Kd[lambda]) at respective wavelengths observed suggests that the spectral composition of the water column changes throughout the year in this region. A significant relationship was observed between Kd(lambda) and aCDOM at 305, 320, 340 and 380 nm only (P < 0.01) but not for 450 nm (PAR) indicating the role of CDOM in regulating variations in Kd(lambda), particularly in the UVR range. The slope S, obtained from a natural-log plot of the absorption coefficient of CDOM against wavelength, ranged between 0.014 and 0.036 nm-1 annually (average = 0.020 +/- 0.007, C.V. = 35%) and suggests seasonal changes in the origin of CDOM between terrestrial (low S) and biogenous (high S) CDOM.

Journal Article↗

PubMed2000

Increased cholesterol absorption by hyperlipidemia atherosclerosis prone (LAP) Japanese quail.

The present study describes the cholesterol absorption by hyperlipidemia atherosclerosis prone (LAP) Japanese quail to address their high susceptibility to experimental atherosclerosis. The apparent cholesterol absorption rate of LAP quail was compared with that of commercially available (CA) Japanese quail. After 14 d of cholesterol feeding by gavage, it was found that the cholesterol excretion of LAP quail was significantly lower than that of CA quail. The fecal excretion of bile acid and fat showed a similar tendency to that as shown with the case of cholesterol. The cholesterol feeding only increased the serum cholesterol level of LAP quail, and this trend holds true for the liver lipid concentration. The expression level of liver cholesterol 7alpha-hydroxylase mRNA showed no difference between LAP and CA strains under the conditions of cholesterol loading. These results showed that the cholesterol absorption by LAP quail is significantly higher than that by CA quail, which may reasonably explain the higher susceptibility of this strain to experimental atherosclerosis.

Animals↗

PubMed2000

A decline in lower extremity lean body mass per body weight is characteristic of women with early phase osteoarthritis of the knee.

OBJECTIVE: Sarcopenia progresses with aging, but the effect of muscle loss on degenerative joint disorders has not been precisely evaluated. We assessed the distribution of lean body mass (LBM) in middle aged women who had osteoarthritis (OA) of the knee for < 5 years. METHODS: Two hundred thirty-five Japanese women aged 45-69 years, including 117 with knee OA for < 5 years, and 118 age and sex matched healthy controls were studied. Body composition measurements were carried out by segmental bioelectrical impedance using 8 surface electrodes. Anthropometric data consisting of LBM of upper extremities/body weight (U-LBM/W), LBM of trunk/body weight (T-LBM/W), and LBM of lower extremities/body weight (L-LBM/W) for knee OA participants were compared to corresponding data for controls. In the knee OA group, the significance of correlations between anthropometric data and Lequesne index of severity for knee OA and between anthropometric data and disease duration were assessed. RESULTS: L-LBM/W was significantly lower in knee OA participants compared with controls (19.2 +/- 2.7% vs 21.0 +/- 2.9%; p < 0.0001). However, no significant differences in U-LBM/W and T-LBM/W were observed between the knee OA and control groups (p > 0.2). L-LBM/W did not correlate significantly with the index of severity or disease duration (p > 0.2). CONCLUSION: Decline in L-LBM/W, but not U-LBM/W or T-LBM/W, may be important in the pathogenesis of knee OA.

Adipose Tissue↗

PubMed2000

[Fukuyama-type congenital muscular dystrophy].

Fukuyama-type congenital muscular dystrophy (FCMD) is characterized by congenital muscular dystrophy in combination with cortical dysgenesis and ocular abnormality. We identified on chromosome 9 q31 the gene for FCMD, which encodes a novel 461-amino-acid protein (fukutin). Most FCMD-bearing chromosomes have been derived from a single ancestral founder, whose mutation consisted of a 3-kb retrotransposal insertion in the 3' non-coding region of the fukutin gene. Some point mutations causing premature termination were found. Amino acid sequence and transfection experiments suggest that fukutin may be an extracellular protein. Pathological study on the brain of the FCMD fetuses revealed that the glia-limitans and basement-membrane complex had frequent breaks. Because of this, developing neurons were shown to overmigrate in the cerebrum. Electron microscopy of the skeletal muscle in FCMD showed that the basal lamina has a disrupted appearance. Thus, a structural alteration of the basal lamina appears to play a key role in the pathophysiology of FCMD. The spectrum of clinical variability of FCMD is much wider than recognized previously. Point mutations have been seen to render the FCMD phenotype rather severe. FCMD could give rise to only in the Japanese who have a milder retrotransposon mutation.

Basement Membrane↗

PubMed1999

Topoisomerase III is essential for accurate nuclear division in Schizosaccharomyces pombe.

Topoisomerases catalyse changes in the topological state of DNA and are required for many aspects of DNA metabolism. While the functions of topoisomerases I and II in eukaryotes are well established, the role of topoisomerase III remains poorly defined. We have identified a gene in the fission yeast Schizosaccharomyces pombe, designated top3 (+), which shows significant sequence similarity to genes encoding topoisomerase III enzymes in other eukaryotic species. In common with murine TOP3 alpha, but in contrast to Saccharomyces cerevisiae TOP3, the S.pombe top3 (+)gene is essential for long-term cell viability. Fission yeast haploid spores containing a disrupted top3 (+)gene germinate successfully, but then undergo only a limited number of cell divisions. Analysis of these top3 mutants revealed evidence of aberrant mitotic chromosome segregation, including the 'cut' phenotype, where septation is completed prior to nuclear division. Consistent with the existence of an intimate association (originally identified in S.cerevisiae ) between topoisomerase III and DNA helicases of the RecQ family, deletion of the rqh1 (+)gene encoding the only known RecQ helicase in S.pombe suppresses lethality in top3 mutants. This conservation of genetic interaction between two widely diverged yeasts suggests that the RecQ family helicases encoded by the Bloom's and Werner's syndrome genes are likely to act in concert with topoisomerase III isozymes in human cells. Our data are consistent with a model in which the association of a RecQ helicase and topoisomerase III is important for facilitating decatenation of late stage replicons to permit faithful chromosome segregation during anaphase.

Amino Acid Sequence↗

PubMed1999

Two distinct ubiquitin-proteolysis pathways in the fission yeast cell cycle.

The SCF complex (Skp1-Cullin-1-F-box) and the APC/cyclosome (anaphase-promoting complex) are two ubiquitin ligases that play a crucial role in eukaryotic cell cycle control. In fission yeast F-box/WD-repeat proteins Pop1 and Pop2, components of SCF are required for cell-cycle-dependent degradation of the cyclin-dependent kinase (CDK) inhibitor Rum1 and the S-phase regulator Cdc18. Accumulation of these proteins in pop1 and pop2 mutants leads to re-replication and defects in sexual differentiation. Despite structural and functional similarities, Pop1 and Pop2 are not redundant homologues. Instead, these two proteins form heterodimers as well as homodimers, such that three distinct complexes, namely SCFPop1/Pop1, SCFPop1/Pop2 and SCFPop2/Pop2, appear to exist in the cell. The APC/cyclosome is responsible for inactivation of CDK/cyclins through the degradation of B-type cyclins. We have identified two novel components or regulators of this complex, called Apc10 and Ste9, which are evolutionarily highly conserved. Apc10 (and Ste9), together with Rum1, are required for the establishment of and progression through the G1 phase in fission yeast. We propose that dual downregulation of CDK, one via the APC/cyclosome and the other via the CDK inhibitor, is a universal mechanism that is used to arrest the cell cycle at G1.

Anaphase-Promoting Complex-Cyclosome↗

PubMed1999

The MAPK kinase Pek1 acts as a phosphorylation-dependent molecular switch.

The mitogen-activated protein kinase (MAPK) pathway is a highly conserved eukaryotic signalling cascade that converts extracellular signals into various outputs, such as cell growth and differentiation. MAPK is phosphorylated and activated by a specific MAPK kinase (MAPKK): MAPKK is therefore considered to be an activating regulator of MAPK. Pmk1 is a MAPK that regulates cell integrity and which, with calcineurin phosphatase, antagonizes chloride homeostasis in fission yeast. We have now identified Pek1, a MAPKK for Pmk1 MAPK. We show here that Pek1, in its unphosphorylated form, acts as a potent negative regulator of Pmk1 MAPK signalling. Mkh1, an upstream MAPKK kinase (MAPKKK), converts Pek1 from being an inhibitor to an activator. Our results indicate that Pek1 has a dual stimulatory and inhibitory function which depends on its phosphorylation state. This switch-like mechanism could contribute to the all-or-none physiological response mediated by the MAPK signalling pathway.

Amino Acid Sequence↗

PubMed1999

A novel nuclear export signal sensitive to oxidative stress in the fission yeast transcription factor Pap1.

Pap1, a fission yeast AP-1-like transcription factor, is negatively regulated by CRM1/exportin 1, the nuclear export factor. Pap1 was localized normally in the cytoplasm but was accumulated in the nucleus when Crm1 was inactivated by a temperature-sensitive mutation or by treatment with leptomycin B, a specific export inhibitor. Deletion of the C-terminal cysteine-rich domain (CRD) resulted in nuclear accumulation of Pap1, while a glutathione S-transferase-green fluorescent protein-CRD fusion protein was localized in the cytoplasm in a Crm1-dependent manner. Deletion and mutational analyses identified several important amino acids in a 19-amino acid region in the CRD as a nuclear export signal (NES). Strikingly, a cysteine residue (Cys-532), in addition to two leucines and an isoleucine, was important for the NES function and the presence of at least one of the two cysteine residues was essential. Unlike classical NESs such as the human immunodeficiency virus Rev NES, the Pap1 NES lost the function upon treatment with oxidants such as diethyl maleate. The oxidative stress response is conserved through evolution, as green fluorescent protein-fused proteins bearing the Pap1 NES expressed in mammalian cells responded to diethyl maleate. These results show that the hydrophobic amino acid-rich region containing two important cysteines in Pap1 serves as a novel NES, which is sensitive to oxidative stress.

Amino Acid Sequence↗