The associations of eating behavior and dietary intake with metabolic syndrome in Japanese: Saku cohort baseline study.

BACKGROUND: The prevention of metabolic syndrome (MetS) is a major public health concern in Japan. The effects of the relationship between eating behavior and nutritional intake on MetS remained unclear. To evaluate nutrition's role in preventing or exacerbating MetS, we examined the associations among eating behavior, nutritional intake, and MetS for the baseline study in the cohort subjects undergone health checkups. METHODS: Four thousand and four hundred forty-seven Japanese men and women were enrolled at the Saku Central Hospital. They received an anthropometric and clinical examination and were assessed for present illness, lifestyle factors such as physical activity, smoking, drinking, and dietary habits at the enrollment. Eating behavior was analyzed by the Sakata's Eating Behavior Questionnaire. Dietary assessment was made using a brief self-administered diet history questionnaire. Two thousand and six hundred two men and 1844 women aged more than 20 were analyzed. RESULTS: The mean age in men and women were 59.2 and 58.4 years old and the mean body mass index (BMI) were 23.7 and 22.3 kg/m2, respectively. The percentages of MetS were 20.6 in men and 6.1 in women. In some nutrients, significantly higher energy-adjusted intakes in subjects without MetS than with Mets appeared both in men and women after age adjustment. After adjusting by age, energy-adjusted intake beverages in men and cereals in women were significantly higher in subjects with MetS than those without MetS. The scores of all the categories in eating behavior were significantly worse in subjects with MetS than those without MetS. CONCLUSIONS: The differences in dietary intake between subjects with Mets and without Mets were relatively small. The scores of all the categories in eating behavior were worse in subjects with MetS than without MetS. It was suggested that the problem lay in the quality of diet, not in the quantity, caused by bad eating habits. The potential influence of eating behavior and nutritional intake on MetS was presented in men and women.

Energy Expenditure in Free-Living Japanese People with Obesity and Type 2 Diabetes, Measured Using the Doubly-Labeled Water Method.

We determined the total energy expenditure (TEE) of healthy overweight or obese people, and those with impaired glucose tolerance and/or impaired fasting glycemia (IGT/IFG), or type 2 diabetes (T2DM) using the doubly-labeled water method. As a second purpose, we compared the measured TEE with the target energy intake recommended in the treatment guidelines for diabetes. The participants were normal glucose tolerance (NGT), and IGT/IFG (n=11) and T2DM (n=9) patients, who were 50-59 y and had a body mass index >25 kg/m2. The median TEE/body mass (BM) values were 32.6, 33.3, and 34.4 kcal/kg BM and the TEE/target BM values (target BM: BM at a BMI of 22 kg/m2) were 43.7, 50.2, and 46.5 kcal/kg target BM for each group, respectively, and did not differ significantly among them. Obese Japanese participants with T2DM in this study had lower TEE/BM than previously studied in non-obese participants with T2DM. In IGT/IFG or T2DM patients, if 30 kcal/kg target BM was used as the energy coefficient, on the basis of the treatment guidelines, the difference between TEE and the target energy intake would be -1,174±552 kcal (-38±11%). When 35 kcal/kg target BM was used as the energy coefficient, the difference between TEE and the target energy intake would be -877±542 kcal (-27±13%). Thus, the energy coefficients used to estimate target energy intake during lifestyle modification in obese/overweight patients with T2DM are considered to be quite low during the first step of diet therapy.

Vegetable Intake at Breakfast and Associated Factors among Young Adults in Japan.

In Japan, the amount of vegetable intake in young adults, especially at breakfast, is low. We aimed to examine the factors associated with vegetable intake at breakfast among young adults. In 2016, a self-administered questionnaire survey was conducted at a Japanese university. The questionnaire measured the frequency of ≥70 g vegetable intake at breakfast, lifestyle, eating habits, environmental factors, knowledge and attitudes (including the variable "Perceived behavior control (PBC)", from the Theory of Planned Behavior). The data of 1,455 male students who usually ate breakfast were analyzed. They were divided into two groups based on the median of the frequency of vegetable intake and the above factors were compared using a chi-squared test by living condition. The significant factors were subsequently entered into a logistic regression analysis as independent variables. Eating breakfast with family or friends and PBC were positively associated with vegetable intake in students living with their family. Often doing one's own cooking, having knowledge about a well-balanced diet, and PBC were positively associated with vegetable intake in students living alone. Along with PBC, different factors pertaining to an individual's living condition may need to be considered to increase vegetable intake at breakfast.

Decreased number of days of fever detection and duration of fever with continuous intake of a fermented milk drink: a randomized, double-blind, placebo-controlled study of elderly nursing home residents.

We conducted a randomized, double-blind, placebo-controlled parallel study to investigate the effects of a fermented milk on elderly nursing home residents. Eighty-eight participants each drank one bottle of fermented milk containing Lactobacillus casei strain Shirota, or a placebo, on a daily basis for 6 months in winter. Peripheral blood, saliva, fecal samples, and clinical data were analyzed to assess the milk's efficacy. Fermented milk consumption was associated with a significant decrease in the number of days on which fever was detected and the mean duration of fever compared with these values in the placebo group. No significant differences were observed in other biological parameters. Continuous intake of this fermented milk could be beneficial for the elderly in terms of suppressing the number of days of detection of fever and the duration of fever, which usually increase in winter.

Effects of 1-year weight loss intervention on abdominal skeletal muscle mass in Japanese overweight men and women.

BACKGROUND AND OBJECTIVES: Limited information is available on how weight loss intervention programs affect skeletal muscle mass especially in trunk. METHODS AND STUDY DESIGN: A total of 235 overweight Japanese men and women aged 40-64 years with a body mass index of 28.0 to 44.8 kg/m2 participated in this randomized controlled intervention study. They were randomly divided into a lifestyle intervention group and control group. Before and after the one-year lifestyle intervention for weight loss an abdominal transverse image was acquired by computed tomography. The cross-sectional areas (CSAs) of visceral fat, subcutaneous fat, and skeletal muscle of rectus abdominis, abdominal oblique, iliopsoas, and erector spinae muscle were calculated. RESULTS: The body weight changed by approximately -5% in the intervention groups. The corresponding values for subcutaneous fat and visceral fat CSAs were -10.8 to -17.5% in both sexes. The reductions observed in skeletal muscle CSAs were significantly less (-6.0% and -7.2% in the men and women intervention groups respectively) than those in fat tissue CSAs. The CSA of each of the four skeletal muscle groups also significantly decreased; however, after adjustments for body weight at each time point, only reductions in the iliopsoas muscle in both sex and abdominal oblique muscles in men remained significant. CONCLUSIONS: The lifestyle weight loss intervention might reduce the relative amount of the abdominal skeletal muscles especially in iliopsoas muscle.

Association of visceral fat area with abdominal skeletal muscle distribution in overweight Japanese adults.

BACKGROUND: Quantitative evaluation of visceral fat mass and skeletal muscle mass is important for health promotion. Recently, some studies suggested the existence of adipocyte-myocyte negative crosstalk. If so, abdominal skeletal muscles may easily and negatively affected not only by the age but also the visceral fat because age-related reduction in abdominal region is greater compared with limbs. OBJECTIVE: We cross-sectionally examined the existence of quantitative associations between visceral fat area and abdominal skeletal muscle distribution in overweight people. METHODS: A total of 230 Japanese males and females who aged 40-64 years and whose body mass index (BMI) was 28.0-44.8kg/m2 participated in this study. The cross-sectional area (CSA) of the visceral fat, subcutaneous fat, and abdominal skeletal muscles, namely, the rectus abdominis, abdominal oblique, erector spinae, and iliopsoas muscles were measured by the computed tomography images. RESULTS: Stepwise regression analyses revealed the existence of sex difference in the relation between visceral fat CSA and other morphological variables. In males, BMI was a positive, and the iliopsoas muscle group CSA was a negative contributor of the visceral fat CSA. In females, both age and BMI were selected as positive contributors. CONCLUSION: These data suggested that the visceral fat CSA may negatively associated with iliopsoas muscle group CSA in males. In females, the visceral fat CSA was not significantly related to the distribution of the abdominal skeletal muscle groups.

Cross-sectional association of the number of neighborhood facilities assessed using postal code with objectively measured physical activity: the Saku cohort study.

Objectives The aim of this study was to examine the association between the number of neighborhood facilities that were assessed according to postal code and objectively measured physical activity by using an accelerometer in community-dwelling Japanese people.Methods The participants included 1,274 Japanese people aged 30-84 years from the Saku cohort study. As neighborhood facilities related to physical activity, we extracted information regarding train stations, supermarkets/convenience stores, postal offices/banks, hospitals/clinics, public offices/community centers, cultural facilities/public children's houses, parks, and sports facilities by using each participant's postal code from the online version of the iTownPages directory published by Nippon Telegraph and Telephone Corporation (NTT) and the official homepage of the Saku City Government Office. We measured each participant's physical activity level using an accelerometer, and calculated the average daily step count and the average weekly period of moderate-to-vigorous intensity (≥3 metabolic equivalents of tasks [METs]) physical activity. The association between two selected physical activity-related variables and the numbers of eight types of neighborhood facilities were analyzed by multivariate logistic regression analysis for people aged 30-64 years and for those aged over 65 years.Results On multivariate logistic regression analysis, meeting the 23 METs h/week of moderate-to-vigorous intensity physical activity was significantly and positively associated with the number of supermarkets/convenience stores in the neighborhood in both age groups. In addition, meeting the desired daily step count outlined in the Japanese National Health Promotion guidelines was positively related to the number of postal offices/banks for people aged over 65 years.Conclusion The results of this study suggest that a sufficient number of neighborhood facilities (i.e., stores, banks, and postal offices) is closely associated with the promotion of physical activity.

Perceptions and practices of commensality and solo-eating among Korean and Japanese university students: A cross-cultural analysis.

BACKGROUND/OBJECTIVES: Commensality, eating together with others, is a major representation of human sociality. In recent time, environments around commensality have changed significantly due to rapid social changes, and the decline of commensality is perceived as a serious concern in many modern societies. This study employs a cross-cultural analysis of university students in two East Asian countries, and examines cross-cultural variations of perceptions and actual practices of commensality and solo-eating. SUBJECTS/METHODS: The analysis was drawn from a free-list survey and a self-administrative questionnaires of university students in urban Korea and Japan. The free-listing survey was conducted with a small cohort to explore common images and meanings of commensality and solo-eating. The self-administrative questionnaire was developed based on the result of the free-list survey, and conducted with a larger cohort to examine reasons and problems of practices and associated behaviors and food intake. RESULTS: We found that Korean subjects tended to show stronger associations between solo-eating and negative emotions while the Japanese subjects expressed mixed emotions towards the practice of solo-eating. In the questionnaire, more Korean students reported they prefer commensality and tend to eat more quantities when they eat commensally. In contrast, more Japanese reported that they do not have preference on commensality and there is no notable difference in food quantities when they eat commensally and alone. Compared to the general Korean cohort finding, more proportion of overweight and obese groups of Korean subjects reported that they tend to eat more when they are alone than normal and underweight groups. This difference was not found in the overweight Japanese subjects. CONCLUSION: Our study revealed cross-cultural variations of perceptions and practices of commensality and solo-eating in a non-western setting.

The relationship of body composition to daily physical activity in free-living Japanese adult men.

The objective of the present study was to investigate whether a previously reported apparent negative relationship between fat mass and daily physical activity in Japanese adult women would also be observed in Japanese adult men. The subjects were grouped into quartiles of BMI and body fat percentage (%BF). The number of steps walked each day and the duration of light- to vigorous-intensity physical activity were assessed by an accelerometer over the same period of time as for the doubly labelled water experiment. The results showed that BMI negatively correlated with the number of steps and time spent in moderate-intensity physical activity, whereas %BF showed a negative relationship with physical activity-related energy expenditure (PAEE)/body weight (BW) and physical activity level. The analysis of data using %BF quartiles revealed that PAEE/BW decreased from the second quartile in which the BMI was < 25 kg/m2. These observations are similar to those reported in our previous study in Japanese adult women. These cross-sectional studies cannot prove causality, and that obesity causes physical inactivity may be the case. However, the results of the present study provide information regarding which physical activity variables should be used in longitudinal studies.

Low-molecular-weight adiponectin and high-molecular-weight adiponectin levels in relation to diabetes.

OBJECTIVE: To evaluate the association between adiponectin complexes (high-molecular-weight [HMW], middle-molecular-weight [MMW], and low-molecular-weight [LMW] adiponectin) and diabetes. DESIGN AND METHODS: We conducted a case-control study, based on a cohort in Saku, Japan. Among 2565 participants, 300 participants with diabetes and 300 matched controls (430 men and 170 women) were analyzed. RESULTS: After adjusting for age, physical activity, hypertension, family history, alcohol use, smoking, and menopausal status, total, HMW, and LMW, but not MMW adiponectin levels were inversely associated with diabetes: total adiponectin, odds ratio comparing the highest with the lowest quartiles, 0.46 (95% confidence interval, 0.25-0.82; P for trend = 0.046); HMW, 0.40 (95%CI, 0.22-0.72; P = 0.046); MMW, 1.04 (95%CI, 0.60-1.77; P = 0.81); and LMW, 0.51 (95%CI, 0.29-0.89; P = 0.01). The associations between total and HMW adiponectin and diabetes attenuated after adjustment for BMI (P = 0.15 and 0.13, respectively), but LMW remained (P = 0.04). When stratified by sex, LMW adiponectin levels were associated with diabetes in men only. None of the associations were significant after adjustment for HOMA-IR. CONCLUSIONS: Decreased LMW, total, and HMW adiponectin levels are associated with diabetes. These associations may be secondary to adiposity or insulin resistance.

Validity of diabetes self-reports in the Saku diabetes study.

BACKGROUND: Diabetes is an important risk factor for cardiovascular disease, certain types of cancer, and death, and self-reports are one of the most convenient methods for ascertaining diabetes status. We evaluated the validity of diabetes self-reports among Japanese who participated in a health checkup. METHODS: Self-reported diabetes was cross-sectionally compared with confirmed diabetes among 2535 participants aged 28 to 85 years in the Saku cohort study. Confirmed diabetes was defined as the presence of at least 1 of the following: fasting plasma glucose (FPG) level of 126 mg/dL or higher, 2-hour post-load glucose (2-hPG) level of 200 mg/dL or higher after a 75-gram oral glucose tolerance test, glycated hemoglobin (HbA1c) level of 6.5% or higher, or treatment with hypoglycemic medication(s). RESULTS: Of the 251 participants with self-reported diabetes, 121 were taking hypoglycemic medication(s) and an additional 69 were classified as having diabetes. Of the 2284 participants who did not self-report diabetes, 80 were classified as having diabetes. These data yielded a sensitivity of 70.4%, a specificity of 97.3%, a positive predictive value of 75.7%, and a negative predictive value of 96.5%. The frequency of participants with undiagnosed diabetes was 3.0%. Of these, 64.2% had FPG within the normal range and were diagnosed by 2-hPG and/or HbA1c. CONCLUSIONS: Our findings provide additional support for the use of self-reported diabetes as a measure of diabetes in epidemiologic studies performed in similar settings in Japan if biomarker-based diagnosis is difficult.

Reduction in adiposity, ß-cell function, insulin sensitivity, and cardiovascular risk factors: a prospective study among Japanese with obesity.

BACKGROUND: A reduction in adiposity may be associated with an improvement in insulin sensitivity and ß-cell function as well as cardiovascular disease (CVD) risk factors; however, few studies have investigated these associations in a longitudinal setting. METHODS: To investigate these associations over a 1-year period, we conducted an observational analysis of 196 Japanese subjects with obesity in the Saku Control Obesity Program. We investigated the relations between changes in adiposity (body mass index [BMI], waist circumference, subcutaneous fat area [SFAT], and visceral fat area [VFAT]) and changes in HbA1c, fasting plasma glucose (FPG), insulin sensitivity index (ISI), the homeostasis model assessment ß cell function (HOMA-ß), lipids, and blood pressure. RESULTS: All adiposity changes were positively associated with HbA1c and FPG changes. Reductions in BMI and VFAT were associated with HOMA-ß reduction. Reductions in all adiposity measures were associated with an improvement in the ISI. Changes in most adiposity measures were positively associated with changes in blood pressure and lipid levels, except for LDL. CONCLUSION: The present findings provide additional supportive evidence indicating that a reduction in adiposity may lead to an improvement in insulin sensitivity and the reduction of CVD risk factors in obese individuals.

Preproghrelin gene polymorphisms in obese Japanese women. Minor homozygotes are light eaters, do not prefer protein or fat, and apparently have a poor appetite.

Preproghrelin gene single-nucleotide polymorphisms are possible predisposing factors to obesity and other metabolic syndromes. To study the correlation between genotypes and obesity, we recruited 117 obese Japanese women (BMI, 25.0-41.1; average, 31.1). Minor homozygotes for five preproghrelin gene polymorphisms, namely, -1500C>G (rs3755777), -1062G>C (rs26311), -994C>T (rs26312) (promoter region), Leu72Met (rs696217) (exon 2), and +3056T>C (rs2075356) (intron 2), had high values of total and visceral fat areas, waist circumference, and BMI, indicating significant correlation of the polymorphisms with obesity and fat metabolism. Here, we studied the relationship between the genotypes and dietary tendency. Self-administered Diet History Questionnaire showed that total food intake, sugar, and dairy product intake were low in +3056C/C women. Their energy, protein, fat, and meat intake was also low. Energy balance calculation showed considerably reduced fat and protein consumption. Dietary habits were surveyed using Sakata's Questionnaire on Eating Behavior. Of the genotypes, -1062C/C women showed low scores for "motivation for eating" and "eating because of stress or something else." Thus, surprisingly, it was revealed that minor homozygotes for preproghrelin gene polymorphisms were light eaters, did not prefer fat or protein, and apparently had a poor appetite, although they were predisposed to obesity.

Associations of sex hormone-binding globulin and testosterone with diabetes among men and women (the Saku Diabetes study): a case control study.

BACKGROUND: Sex hormone-binding globulin (SHBG) levels and sex hormones have been implicated in the pathogenesis of type 2 diabetes and cardiovascular diseases. As fatty liver has been suggested to be a major determinant of SHBG levels, we examined whether the associations of SHBG and testosterone with diabetes were independent of fatty liver. METHODS: We conducted a case-control study that included 300 diabetes cases (215 men and 85 women) and 300 matched controls from the Saku cohort study. Diabetes was defined by either fasting plasma glucose levels ≥126 mg/dL, 2-h post-load glucose levels ≥200 mg/dL after a 75 g oral glucose tolerance test, or diabetes diagnosed by physicians. We fitted conditional logistic regression models to examine the associations between SHBG and total testosterone levels with diabetes by sex. To evaluate the impact of fatty liver, we used the fatty liver index (FLI), a validated measure derived from serum triglyceride levels, body mass index (BMI), waist circumference, and γ-glutamyltransferase levels. RESULTS: After adjusting for age, family history of diabetes, smoking, physical activity, BMI, and FLI, SHBG levels were inversely associated with diabetes among women (odds ratio [OR] comparing the highest with the lowest quartiles, 0.13 [95% confidence interval {CI}, 0.02-0.96]), but not among men. Similar patterns were observed in a subgroup analysis restricted to postmenopausal women"(OR, 0.12 [95% CI, 0.01-1.17]). In contrast, testosterone levels were inversely associated with diabetes among men (OR, 0.45 [95% CI, 0.23-0.89]), but not among women. CONCLUSIONS: Our findings suggest that SHBG in women and testosterone in men may be inversely associated with diabetes.

Dietary glycemic index and glycemic load in relation to HbA1c in Japanese obese adults: a cross-sectional analysis of the Saku Control Obesity Program.

BACKGROUND: Dietary glycemic index or load is thought to play an important role in glucose metabolism. However, few studies have investigated the relation between glycemic index (GI) or load (GL) and glycemia in Asian populations. In this cross-sectional analysis of a randomized controlled trial, the Saku Control Obesity Program, we examined the relation between the baseline GI or GL and glycemia (HbA1c and fasting plasma glucose [FPG] levels), insulin resistance (HOMA-IR), ß-cell function (HOMA-ß), and other metabolic risk factors (lipid levels, diastolic and systolic blood pressure, and adiposity measures). METHODS: The participants were 227 obese Japanese women and men. We used multiple linear regression models and logistic regression models to adjust for potential confounding factors such as age, sex, visceral fat area, total energy intake, and physical activity levels. RESULTS: After adjustments for potential confounding factors, GI was not associated with HbA1c, but GL was positively associated with HbA1c. For increasing quartiles of GI, the adjusted mean HbA1c were 6.3%, 6.7%, 6.4%, and 6.4% (P for trend = 0.991). For increasing quartiles of GL, the adjusted mean HbA1c were 6.2%, 6.2%, 6.6%, and 6.5% (P for trend = 0.044). In addition, among participants with HbA1c ≥ 7.0%, 20 out of 28 (71%) had a high GL (≥ median); the adjusted odds ratio for HbA1c ≥ 7.0% among participants with higher GL was 3.1 (95% confidence interval [CI] = 1.2 to 8.1) compared to the participants with a lower GL (

What behaviors are important for successful weight maintenance?

Purpose. To examine behavioral factors related to successful weight maintenance. Methods. Subjects were 90 middle-aged participants who attended a weight loss program and were followed for one year. The subjects were classified into either successful weight maintainers (maintained a weight loss of 5% or more from their initial weight for one year) (SWM) or unsuccessful weight maintainers (USWM), and weight control practice, stress, obstacles, support, and self-efficacy during the program and follow-up period were compared. Results. SWM had mean loss of 12% from their initial weight during the program. They showed a greater improvement in their regularity of eating, walked more, and felt less stress regarding their increased physical activity than the USWM. During the follow-up period, significantly more SWM participants had self-efficacy (for measuring weight, practicing dietary objective, and assessing the practice and keeping records), actually kept records and measured weight more than the USWM participants. In contrast, more USWM participants felt stress about measuring weight. Conclusion. In addition to a substantial initial weight loss due to an increased amount of physical activity, having a higher self-efficacy and consistently keeping records of one's activities, as well as regularly weighing themselves, may be important for successful weight maintenance.

Behavioral change during weight loss program and one-year follow-up: Saku Control Obesity Program (SCOP) in Japan.

This study evaluated effects of a behavioral approach which placed emphasis on tailored behavior counseling, diet, weight loss and weight maintenance. A one-year randomized controlled trial was conducted among 235 overweight/obese adults in Japan. The intervention group (n=119) received individual-based counseling using a behavioral approach and the changes made in the diet and physical activity were dependent on each participant as much as possible. One year later, the intervention group lost significantly more weight than the control group (-5.0 kg vs. 0.1 kg for men and -3.9 kg vs. -0.2 kg for women). Compared to the control group, the male intervention group reduced overall energy, cereals and dairy products consumption significantly, while increasing green and yellow vegetable intake, and the female intervention group significantly reduced intake of dairy products. Regarding behaviors, both male and female intervention groups increased the number of walking steps and women improved their irregular eating habits compared to those in the control groups. Behavior changes were related to weight loss; participants who maintained the action/maintenance stage or moved to later stages lost significantly more weight than participants who remained in the pre-contemplation/contemplation/preparation stages or regressed to earlier stages. After one-year follow-up, the intervention group maintained significantly lower weights, lower energy intakes and improvements in irregular eating habits. Our behavioral approach led to diet and behavior modification, weight loss and maintenance. Because modified variables differed between men and women, gender-specific approaches may be necessary.

Relation of body composition to daily physical activity in free-living Japanese adult women.

The objective of the present study was to investigate the relationship between the indices of body size such as BMI, fat-free mass index (FFMI, FFM/height2), fat mass index (FMI, FM/height2), and body fat percentage (%BF), and physical activities assessed by the doubly-labelled water (DLW) method and an accelerometer in free-living Japanese adult women. We conducted a cross-sectional study in 100 female subjects ranging in age from 31 to 69 years. Subjects were classified in quartiles of BMI, FFMI, FMI and %BF. Daily walking steps and the duration of light to vigorous physical activity were simultaneously assessed by an accelerometer for the same period as the DLW experiment. Only physical activity-related energy expenditure (PAEE)/FFM and PAEE/body weight (BW) decreased in the highest quartile of BMI. Physical activity level, PAEE/FFM and PAEE/BW decreased in the highest quartile of FMI and %BF, whereas they were not different among quartiles of FFMI. Daily walking steps and the duration of moderate- and vigorous-intensity physical activities decreased or tended to decrease in the highest quartile of FMI and %BF, but did not differ among quartiles of FFMI and BMI. These results clearly showed that Japanese adult women with higher fat deposition obviously had a low level of physical activities assessed by both the DLW method and accelerometry, but those with larger BMI had lower PAEE/FFM and PAEE/BW only. Our data suggest that the relationship between obesity and daily physical activities should be discussed using not only BMI but also FMI or %BF.

Caffeine abolishes the ultraviolet-induced REV3 translesion replication pathway in mouse cells.

When a replicative DNA polymerase stalls upon encountering a photoproduct on the template strand, it is relieved by other low-processivity polymerase(s), which insert nucleotide(s) opposite the lesion. Using an alkaline sucrose density gradient sedimentation technique, we previously classified this process termed UV-induced translesion replication (UV-TLS) into two types. In human cancer cells or xeroderma pigmentosum variant (XP-V) cells, UV-TLS was inhibited by caffeine or proteasome inhibitors. However, in normal human cells, the process was insensitive to these reagents. Reportedly, in yeast or mammalian cells, REV3 protein (a catalytic subunit of DNA polymerase ζ) is predominantly involved in the former type of TLS. Here, we studied UV-TLS in fibroblasts derived from the Rev3-knockout mouse embryo (Rev3KO-MEF). In the wild-type MEF, UV-TLS was slow (similar to that of human cancer cells or XP-V cells), and was abolished by caffeine or MG-262. In 2 cell lines of Rev3KO-MEF (Rev3(-/-)p53(-/-)), UV-TLS was not observed. In p53KO-MEF, which is a strict control for Rev3KO-MEF, the UV-TLS response was similar to that of the wild-type. Introduction of the Rev3 expression plasmid into Rev3KO-MEF restored the UV-TLS response in selected stable transformants. In some transformants, viability to UV was the same as that in the wild-type, and the death rate was increased by caffeine. Our findings indicate that REV3 is predominantly involved in UV-TLS in mouse cells, and that the REV3 translesion pathway is suppressed by caffeine or proteasome inhibitors.

Rev1, Rev3, or Rev7 siRNA Abolishes Ultraviolet Light-Induced Translesion Replication in HeLa Cells: A Comprehensive Study Using Alkaline Sucrose Density Gradient Sedimentation.

When a replicative DNA polymerase stalls upon encountering a lesion on the template strand, it is relieved by other low-processivity polymerase(s), which insert nucleotide(s) opposite the lesion, extend by a few nucleotides, and dissociate from the 3'-OH. The replicative polymerase then resumes DNA synthesis. This process, termed translesion replication (TLS) or replicative bypass, may involve at least five different polymerases in mammals, although the participating polymerases and their roles have not been entirely characterized. Using siRNAs originally designed and an alkaline sucrose density gradient sedimentation technique, we verified the involvement of several polymerases in ultraviolet (UV) light-induced TLS in HeLa cells. First, siRNAs to Rev3 or Rev7 largely abolished UV-TLS, suggesting that these 2 gene products, which comprise Polζ, play a main role in mutagenic TLS. Second, Rev1-targeted siRNA also abrogated UV-TLS, indicating that Rev1 is also indispensable to mutagenic TLS. Third, Polη-targeted siRNA also prevented TLS to a greater extent than our expectations. Forth, although siRNA to Polι had no detectable effect, that to Polκ delayed UV-TLS. To our knowledge, this is the first study reporting apparent evidence for the participation of Polκ in UV-TLS.