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PubMed2001

Kap promoter analysis in vivo: a regulatory role for a truncated L1 repeat.

Reporter gene expression directed by a 1542-base pair (bp) fragment of the Kap promoter is specific to the proximal tubules of the kidney and androgen-regulated. In the present study, the characteristics of the androgen response from the 1542-bp promoter were examined in vivo. The estrogen response in the kidney and uterus was also examined. The reporter gene expression was assayed in lines of transgenic mice generated from a truncated promoter construct in which the L1 repeat, present at the distal portion of the 1542-bp, had been deleted. The pattern of androgen response of the reporter gene is similar to that of the endogenous Kap. Reporter gene expression in the 1542-bp promoter does not respond to estrogen in the kidney, while perinatal expression in the uterus does occur. Truncation of the L1 results in loss of reporter gene expression. We conclude that L1 sequences present near the Kap promoter have a regulatory function.

Angiotensinogen↗
PubMed1998

Purification and crystallization of the CDK-associated protein phosphatase KAP expressed in Escherichia coli.

The kinase associated phosphatase (KAP) is a human dual specificity protein phosphatase that dephosphorylates the cell cycle control protein, cyclin dependent kinase-2 on Thr 160 in a cyclin dependent manner (Poon & Hunter, 1995). We report here the over-expression of KAP in Escherichia coli as an N-terminal His-tagged protein using a modified pET-28a T7-expression vector. The recombinant protein was purified to homogeneity and crystallized. The crystals diffract to 2.3 A resolution when exposed to synchrotron radiation and belong to space group P6(1)22, or its enantiomorph P6(5)22, with unit cell dimensions a = b = 74.5 A, c = 139.5 A.

Amino Acid Sequence↗
PubMed1996

Effects of thyroid hormone on the androgenic expression of KAP gene in mouse kidney.

The kidney androgen-regulated protein (KAP) gene exhibits a cell-specific hormonal regulation of its expression in the epithelial cells of proximal tubules of mouse kidney, where T3 is required for constitutive expression in the straight segments and androgens for expression in the convoluted ones. By using different models of hypothyroidism, we demonstrate that maximal androgen-mediated induction of the gene depends on thyroid hormone as well. This constitutes a specific event, since vitamin D3 cannot mimic the effects of T3, albeit their remarkable functional relationship. It is also shown that while congenital hypothyroid hyt/hyt male mice, exposed to maternal T3 in the gestational period, exhibit diminished but existent androgen-dependent cortical responses, mice exposed to goitrogens during gestation and postnatally are unable to express the gene even at postnatal day ninety. Impairment of KAP cortical expression in hypothyroid animals does not correlate with lower levels of androgens or androgen receptor expression.

Animals↗
PubMed1999

Pmg-1 and pmg-2 constitute a novel family of KAP genes differentially expressed during skin and mammary gland development.

The epidermis, by invagination of the undifferentiated ectodermal cells, gives rise to several distinct structures including hair, sebaceous, eccrine sweat and mammary glands. We have recently isolated a novel gene, pmg-1, expressed in the pubertal mouse mammary gland. While investigating its genomic structure, we identified a related gene in close proximity, which we have termed pmg-2. pmg-1 and pmg-2 are intron-less, are transcribed in opposite directions and are separated by a potential promoter region of 2.8 kb containing putative binding motifs for the developmental transcription factors Lef-1, Sox5 and D-STAT. pmg-1 and pmg-2 encode small proteins rich in G, S, F, Y and Q and contain characteristic repeats reminiscent of the keratin-associated proteins (KAPs). Both genes are expressed in growing hair follicles in skin as well as in sebaceous and eccrine sweat glands. Interestingly, expression is also detected in the mammary epithelium where it is limited to the onset of the pubertal growth phase and is independent of ovarian hormones. Their broad, developmentally controlled expression pattern, together with their unique amino acid composition, demonstrate that pmg-1 and pmg-2 constitute a novel KAP gene family participating in the differentiation of all epithelial cells forming the epidermal appendages.

Amino Acid Sequence↗
PubMed2003

Activating MAP KAP kinase 2.

The crystal structures of MAP KAP kinase 2 in complex with ADP and staurosporine suggest a major reorganization of the glycine-rich loop upon ligand binding. The main determinant for the catalytic activity of MAP KAP kinase 2 is phosphorylation by p38 MAP kinase.

Adenosine Diphosphate↗
PubMed2000

A novel nuclear receptor corepressor complex, N-CoR, contains components of the mammalian SWI/SNF complex and the corepressor KAP-1.

Transcriptional silencing by many transcription factors is mediated by the nuclear receptor corepressor (N-CoR). The mechanism by which N-CoR represses basal transcription involves the direct or indirect recruitment of histone deacetylases (HDACs). We have isolated two multiprotein N-CoR complexes, designated N-CoR-1 and N-CoR-2, which possess histone deacetylase activity that is mediated by distinct HDACs. Based on Western blotting using antibodies against known subunits, the only HDAC found in the N-CoR-1 complex was HDAC3. In contrast, N-CoR-2 contained predominantly HDAC1 and HDAC2 as well as several other subunits that are found in the Sin3A.HDAC complex. Using mass spectrometry and Western blotting, we have identified several novel components of the N-CoR-1 complex including the SWI/SNF-related proteins BRG1, BAF 170, BAF 155, BAF 47/INI1, and the corepressor KAP-1 that is involved in silencing heterochromatin. Indirect immunofluorescence has revealed that both KAP-1 and N-CoR colocalize throughout the nucleus. These results suggest that N-CoR is found in distinct multiprotein complexes, which are involved in multiple pathways of transcriptional repression.

Blotting, Western↗
PubMed2001

Solution structure of the PHD domain from the KAP-1 corepressor: structural determinants for PHD, RING and LIM zinc-binding domains.

Plant homeodomain (PHD) domains are found in >400 eukaryotic proteins, many of which are transcriptional regulators. Naturally occurring point mutations or deletions of this domain contribute to a variety of human diseases, including ATRX syndrome, myeloid leukemias and autoimmune dysfunction. Here we report the first structural characterization of a PHD domain. Our studies reveal that the PHD domain from KAP-1 corepressor binds zinc in a cross-brace topology between anti-parallel ss-strands reminiscent of RING (really interesting new gene) domains. Using a mutational analysis, we define the structural features required for transcriptional repression by KAP-1 and explain naturally occurring, disease-causing mutations in PHD domains of other proteins. From a comparison of this PHD structure with previously reported RING and LIM (Lin11/Isl-1/Mec-3) structures, we infer sequence determinants that allow discrimination among PHD, RING and LIM motifs.

Amino Acid Sequence↗
PubMed1974

The fractionation of calf thymus DNA by multiple formation of sedimentable complexes with homologous lysine-rich histone KAP. Derivative melting curves and ultracentrifugation in CsCl concentration gradients.

The process of fractionation of total calf thymus DNA using a step precipitation of DNA by means of increasing concentrations of the homologous histone KAP was investigated. In addition to the known fractions three so far undescribed ones/in thymus/,characterized by buoyant densities in CsCl equal 1.692, 1.706 and 1.728 g/ccm, were identified. Considerable amounts of preparations seriously enriched in individual satellite fractions were obtained. The ability of GC-rich satellite DNAs to form more soluble complexes with histone KAP is suggested as reason for the observed fractionation.

Animals↗
PubMed1974

The fractionation of calf thymus DNA on histone KAP-Sepharose 4B columns.

It was found that fractionation of calf thymus DNA on homologous histtone KAP covalently bound to CNBr activated Sepharose 4B depends on the molecular weight of DNA. The maximum of elution of high molecular DNA (m. wt. above 5 x 10(6)) was observed at 0.56 M NaCl and that of degraded DNA (m. wt. 0.8 x 10(6)) at 0.52 M NaCl. Significant differences in melting temperatures and melting intervals were observed among fractions obtained from low molecular DNA as a result of enrichment of some fractions in satellite DNAs. These differences were very small in DNA of m. wt. above 5 x 10(6). The results are discussed in terms of specific areas which may exist on calf thymus DNA molecules playing the role of loci, where lysine-rich histone KAP is preferentially bound.

Animals↗
PubMed2006

Using knowledge, attitudes and practice (KAP) surveys on lymphatic filariasis to prepare a health promotion campaign for mass drug administration in Alor District, Indonesia.

We report the results of two surveys of people's knowledge, attitudes and practices (KAP) regarding lymphatic filariasis (LF) in Alor District, eastern Indonesia. The results of the surveys were used to prepare and evaluate the social mobilization component of a pilot mass drug administration (MDA) in five villages. In the study area, the filarial parasites Brugia timori and Wuchereria bancrofti are highly endemic. Frequent and severe adverse reactions after MDA may occur especially in areas endemic for B. timori and therefore, a special communication strategy was designed to inform and to educate communities about LF and its control. The first KAP survey was conducted as a baseline pre-MDA with diethylcarbamazine and albendazole and the second as a post-intervention evaluation in order to obtain information on the impact of the communication campaign. Before the information campaign and the subsequent MDA, 54% of the study population had heard of one of the three main terms for LF, whereas after health education and MDA, 89% had heard of at least one of the three terms. Similarly, pre-MDA, 21% reported having had previously taken the treatment for filariasis, while post-MDA, 88% reported having taken the treatment during the pilot treatment period. The historical fears and traumatic experiences associated with past LF treatment campaigns in Indonesia were averted since both the communication campaign and the MDA were designed appropriately for and together with the community. As a result, compliance was sufficient in the first round to successfully begin the elimination process.

Adolescent↗
PubMed1994

Knowledge, attitude and practice (KAP) towards epilepsy in secondary school students in Tanzania.

Persons with epilepsy are shunned and discriminated against in education, employment, and marriage in Africa, because epilepsy is seen as a highly contagious and shameful disease in the eyes of the public. These observations come from many studies carried out in Africa including recent ones in Nigeria and Liberia. The KAP towards epilepsy among school children in Tanzania has not been studied and this information is essential before a health education programme on epilepsy can be designed for incorporation into the school curriculum. Therefore a study was carried out on 426 secondary school children in Mahenge to find out their KAP towards epilepsy. Forty six pc (193 of 420, students attributed epilepsy to heredity, brain injury, brain infection and witchcraft in that order. About 60 pc (186 of 331) considered epilepsy to be contagious through physical contact, saliva, and breath. Hospital treatment was considered by 62.7 pc (248 of 395) students and 80 pc (346 of 422) considered epilepsy to be controllable. Over 90 pc (385 of 414) of the students believed an epileptic person to be mentally subnormal and considered this a major reason for not sending an epileptic child to school. Their belief in the contagiousness of epilepsy was a major reason for discriminating against an epileptic child or person playing with other children; going to school; sharing the same plate or bed and as a marriage partner and this was significantly more so in the lower than upper classes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

Knowledge, attitudes and practices (KAP) of human intestinal helminths (worms) in two rural communities in Nyanza Province, Western Kenya.

Six common health problems prevalent in two rural communities-Buoye and Kanyawegi Sublocations--of Kisumu District, in Western Kenya, were selected and the knowledge, attitudes and practices (KAP) of the communities investigated with special reference to human intestinal helminths (worms). The selected diseases were malaria, diarrhoea, human intestinal helminths, scabies, schistosomiasis, and measles. The selection of the six health problems was based on a previous more general KAP study of health and disease at household level in the two areas and information from the Medical Officer of Health in the area. The specific objective was to assess the perceived importance of intestinal worms vis-a-vis other common and known health problems in the community. The study included an analysis of responses to questions on human intestinal worms broken down by sex, age and education level of respondents as well as identification and perceived health effects of a set of vectors and parasites of common diseases in the area. Compared with the other five health problems, intestinal worms did not rank highly in people's minds as an important health problem, although very high proportions of respondents knew of the problem, notably the young and educated. This high proportion of respondents who knew of the problem could describe the symptoms with some accuracy and could correctly identify the vectors and parasite samples. But the full implications of intestinal worms were not fully appreciated. Both communities reflected relatively poor comprehension of causes, treatment and methods of prevention.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1976

Analyst biases in KAP surveys: a cross-cultural comparison.

The analysis phase of the KAP survey, in which meanings are ascribed to found empirical relationships, has largely been ignored in investigations of this data-gathering method. In order to determine the extent to which a lack of familiarity with the culture of data origin might influence survey interpretation, standardized KAP survey data from Bangladesh were presented to four groups of analysts who differed in degree of familiarity with the Bangladesh culture. Meanings ascribed by the analysts to the same empirical relationships were found to vary significantly by the degree to which they were familiar with the culture.

Bangladesh↗
PubMed2005

The Krüppel-like zinc-finger protein ZNF224 represses aldolase A gene transcription by interacting with the KAP-1 co-repressor protein.

Transcription factors belonging to the Krüppel-like zinc finger family of proteins participate in the regulation of cell differentiation and development. Although many of these proteins have been identified, little is known about their structure and function. We recently purified ZNF224, a new Krüppel-like zinc finger protein, that contains a Krüppel-associated box (KRAB) domain at the NH2 terminus, and 19 Cys2-His2 zinc-finger domains at the COOH terminus. Using chromatin immunoprecipitation and transient transfection assays, we demonstrate that ZNF224 binds in vivo to the distal promoter of the aldolase A gene and represses its transcription. The results of transient co-transfection experiments show that ZNF224-mediated transcription repression requires the 45-amino acid long KRAB A domain. The ability of KRAB-containing ZNF224 protein to repress transcription depends on specific interaction with the KAP-1 co-repressor molecule. Finally, using selective treatment with the HDAC1 inhibitor trichostatin A, we demonstrate that ZNF224-mediated repression requires histone deacetylases.

Acetylation↗
PubMed2006

Is vacuum extraction still known, taught and practiced? A worldwide KAP survey.

OBJECTIVE: Is vacuum extraction-the method of first choice for assisting vaginal delivery in case of prolonged labor-losing ground in the developing world? And if it is, why? The paper tries to answer these disturbing questions, and examine their consequences. METHODS: A rapid Knowledge-Attitude-Practice (KAP) survey was conducted during 2003-2004 on the question of assisted vaginal delivery (AVD) by the use of the vacuum extractor. Public health specialists and obstetricians from 121 developing countries were consulted about their knowledge of the method in their country, its reputation (i.e. their attitude) and its use (practice). RESULTS: Overall 48% of the respondent countries have confirmed knowledge, positive attitude, teaching and countrywide use of the method, while 37% said the method is known and used by only a limited number of specialists who do not teach it, and 15% admitted no knowledge and therefore no use. CONCLUSION: Given the evidence-based international recognition of the benefits of vacuum extraction (if practiced correctly and for appropriate indications), it is unjust to deprive women with prolonged labor (and their fetuses) of a simple intervention that can contribute to reducing life threatening complications. This unsophisticated worldwide survey, while not providing in-depth explanations, calls for rehabilitation of vacuum extraction in countries where it is disappearing and surgical extraction is not yet readily accessible to all women with prolonged labor.

Clinical Competence↗
PubMed2003

Catalytically active MAP KAP kinase 2 structures in complex with staurosporine and ADP reveal differences with the autoinhibited enzyme.

MAP KAP kinase 2 (MK2), a Ser/Thr kinase, plays a crucial role in the inflammatory process. We have determined the crystal structures of a catalytically active C-terminal deletion form of human MK2, residues 41-364, in complex with staurosporine at 2.7 A and with ADP at 3.2 A, revealing overall structural similarity with other Ser/Thr kinases. Kinetic analysis reveals that the K(m) for ATP is very similar for MK2 41-364 and p38-activated MK2 41-400. Conversely, the catalytic rate and binding for peptide substrate are dramatically reduced in MK2 41-364. However, phosphorylation of MK2 41-364 by p38 restores the V(max) and K(m) for peptide substrate to values comparable to those seen in p38-activated MK2 41-400, suggesting a mechanism for regulation of enzyme activity.

Adenosine Diphosphate↗
PubMed1995

Chromosomal mapping of the genes for the human cell cycle proteins cyclin C (CCNC), cyclin E (CCNE), p21 (CDKN1) and KAP (CDKN3).

Many gene products associated with the cdk cell cycle kinases are thought to regulate the active kinase complex and thus regulate the transition points of the cell cycle. Genes encoding these proteins may potentially function as oncogenes or tumor suppressor genes. We describe the chromosomal mapping by FISH of the genes for several cdk-associated proteins including human CCNC (cyclin C) to 6q21, CCNE (cyclin E) to 19q12-->q13, CDKN1 (p21) to 6p21.2 and CDKN3 (KAP to 14q22). Some of these sites are near chromosomal translocations or LOH sites common to a variety of human tumors. The potential role for each of these genes in neoplasia is discussed.

Cell Cycle Proteins↗
PubMed1995

Kuopio Atherosclerosis Prevention Study (KAPS). A population-based primary preventive trial of the effect of LDL lowering on atherosclerotic progression in carotid and femoral arteries.

BACKGROUND: The atherosclerotic progression-reducing effect of LDL cholesterol (LDL-C) lowering has been established in subjects with severe atherosclerotic disease but not in persons with elevated LDL cholesterols without severe atherosclerosis. KAPS (Kuopio Atherosclerosis Prevention Study) is the first population-based trial in the primary prevention of carotid and femoral atherosclerosis. METHODS AND RESULTS: The eligibility requirements were serum LDL-C > or = 4.0 mmol/L and total cholesterol < 7.5 mmol/L. Out of a geographically defined population, 447 men aged 44 to 65 years (mean, 57) were randomized to pravastatin (40 mg/d) or placebo for 3 years. Less than 10% of the subjects had prior myocardial infarction. Thirty-nine men discontinued study medication; however, efficacy data were available for 424 men. The primary outcome was the rate of carotid atherosclerotic progression, measured as the linear slope over annual ultrasound examinations in the average of the maximum carotid intima-media thickness (IMT) of the far wall of up to four arterial segments (the right and left distal common carotid artery and the right and left carotid bulb). For the carotid arteries, at the overall mean baseline IMT of 1.66 mm, the rate of progression of carotid atherosclerosis was 45% (95% CI, 16 to 69%) less in the pravastatin (0.017 mm/y) than the placebo (0.031 mm/y) group (P = .005). In the common carotid artery there was a treatment effect of 66% (95% CI, 30 to 95%; pravastatin 0.010 mm/y; placebo 0.029 mm/y; P < .002) at the overall mean baseline IMT of 1.35 mm. A treatment effect of 30% (95% CI, -1% to 54%) was found for the carotid bulb (pravastatin, 0.028; placebo, 0.040; P = .056) at the overall mean baseline IMT of 2.0 mm. The treatment effect was larger in subjects with higher baseline IMT values, in smokers and in those with low plasma vitamin E levels. There was no significant treatment effect on atherosclerotic progression in the femoral arteries. CONCLUSIONS: These data establish the antiatherogenic effect of LDL-C lowering by pravastatin in hypercholesterolemic men in a primary prevention setting and suggest a greater effect in smokers than in nonsmokers.

Anticholesteremic Agents↗