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PubMed2003

Phage HK022 Nun protein represses translation of phage lambda N (transcription termination/translation repression).

The N-terminal arginine-rich motif of phage HK022 Nun protein binds to NUT sequences in phage lambda nascent transcripts and induces transcription termination. Interactions between the Nun C terminus and RNA polymerase as well as the DNA template are required for termination. We have isolated Nun C-terminal point and deletion mutants that are unable to block transcription. The mutants bind NUT RNA and inhibit translation of the lambda N gene. Thus HK022 excludes lambda both by terminating transcription on the phage chromosome and by preventing translation of the essential lambda N gene. Like N autoregulation, translation repression by Nun requires host RNaseIII deficiency (rnc) or a mutation in the RNaseIII processing site (rIII) located between NUTL and the beginning of the N coding sequence. Our data support the idea that Nun bound at NUTL causes steric interference with ribosome attachment to the nearby N coding sequence. Two models, Nun acting alone or in complex with host proteins, are discussed.

Bacteriophage lambda↗
PubMed1993

Use of a gene encoding a suppressor tRNA as a reporter of transcription: analyzing the action of the Nun protein of bacteriophage HK022.

The Nun protein of phage HK022 blocks the expression of genes that lie downstream of the nut sites of phage lambda. Nun is believed to act by promoting premature termination of transcription at or near these sites. To test this hypothesis and to facilitate mapping the sites of termination, we inserted a gene encoding a suppressor tRNA immediately downstream of the lambda nutL site and determined the effect of Nun on tRNA level. We found that Nun severely reduced the accumulation of mature, biologically active tRNA and promoted the accumulation of short, promoter-proximal transcripts whose 3' ends were dispersed over a 100-nucleotide region downstream of nutL. These results are consistent with the hypothesis that Nun terminates transcription within the region immediately downstream of nutL and are inconsistent with the hypothesis that the only action of Nun is to prevent translation of genes located downstream of the nut site. The stability, small size, and easily assayable biological function of suppressor tRNA recommend it as a reporter of transcription in other systems.

Bacteriophage lambda↗
PubMed1995

Interaction between the phage HK022 Nun protein and the nut RNA of phage lambda.

The nun gene product of prophage HK022 excludes phage lambda infection by blocking the expression of genes downstream from the lambda nut sequence. The Nun protein functions both by competing with lambda N transcription-antitermination protein and by actively inducing transcription termination on the lambda chromosome. We demonstrate that Nun binds directly to a stem-loop structure within nut RNA, boxB, which is also the target for the N antiterminator. The two proteins show comparable affinities for boxB and they compete with each other. Their interactions with boxB are similar, as shown by RNase protection experiments, NMR spectroscopy, and analysis of boxB mutants. Each protein binds the 5' strand of the boxB stem and the adjacent loop. The stem does not melt upon the binding of Nun or N, as the 3' strand remains sensitive to a double-strand-specific RNase. The binding of RNA partially protects Nun from proteolysis and changes its NMR spectra. Evidently, although Nun and N bind to the same surface of boxB RNA, their respective complexes interact differently with RNA polymerase, inducing transcription termination or antitermination, respectively.

Bacteriophage lambda↗
PubMed2000

The carboxyl terminus of phage HK022 Nun includes a novel zinc-binding motif and a tryptophan required for transcription termination.

The amino-terminal arginine-rich motif of the phage HK022 Nun protein binds phage lambda nascent mRNA transcripts while the carboxy-terminal domain binds RNA polymerase and arrests transcription. The role of specific residues in the carboxy-terminal domain in transcription termination were investigated by mutagenesis, in vitro and in vivo functional assays, and NMR spectroscopy. Coordination of zinc to three histidine residues in the carboxy-terminus inhibited RNA binding by the amino-terminal domain; however, only two of these histidines were required for transcription arrest. These results suggest that additional zinc-coordinating residues are supplied by RNA polymerase in the context of the Nun-RNA polymerase complex. Substitution of the penultimate carboxy-terminal tryptophan residue with alanine or leucine blocks transcription arrest, whereas a tyrosine substitution is innocuous. Wild-type Nun fails to arrest transcription on single-stranded templates. These results suggest that Nun inhibition of transcription elongation is due in part to interactions between the carboxy-terminal tryptophan of Nun and double-stranded DNA, possibly by intercalation. A model for the termination activity of Nun is developed on the basis of these data.

Amino Acid Sequence↗
PubMedDate not supplied

Growth factor NUN increased AKR1C2 expression by activated CDK2 related RB signal transduction pathway in human liver cancer cell line.

BACKGROUND/AIMS: The investigation is to understand whether polypeptide growth factor NUN can enhance AKR1C2 transcriptional expression in human SMMC7721 liver cancer cell line or not. METHODOLOGY: SMMC7721 cell was starved in serum-free medium overnight before NUN treatment, and cells were then incubated with NUN for different times and dosage. These cells lysates were prepared to examine AKR1C2 expression by Western blot. Total RNA was extracted from these cells to analyze the change of AKR1C2 mRNA by Northern blot. RESULTS: NUN increased expression of AKR1C2 in SMMC7721 liver cancer cell in a time- and dosage-dependent manner and this increase resulted from up-regulation of AKR1C2 mRNA expression. The inhibitor of CDK2 inhibited the up-regulation of AKR1C2 by NUN. CONCLUSIONS: These results showed that NUN can enhance AKR1C2 transcription by activated CDK2 related RB signaling pathway, leading to increased AKR1C2 expression in SMMC7721 liver cancer cell.

CDC2-CDC28 Kinases↗
PubMedDate not supplied

[A window on the past: lay nurses and nun nurses in hospital modernization at the start of the 20th century].

The presence of nuns in Italian hospitals raised hot debates since the beginning of the century. Though recognised by a large majority as ideal nurses, for the devotion and the amount of time spent working in the hospital, physicians, feminist movements and nursing leagues expressed much criticism against nuns, for several reasons: a. nuns were more liable to the Mother Superior than to the hospital administration, therefore could not be neither controlled nor sanctioned; b. misplaced "religious" behaviors and beliefs (i.e. the body as an occasion of sin; suffering means spiritual salvation) which may threaten proper care for patients; c. nuns could not act as educators or role model for nurses because they were mainly involved in organizational and management activities and not in direct patient care; d. among other problems, the way of dressing, with large hats and long sleeves was a source of transport of microorganisms). Lack of properly educated and trained lay nurses, able to substitute nuns, some improvement in nuns' education, strongly promoted by Pope Pio X and the dissolution of nursing leagues put an end to the fight for the secularization of hospitals.

Catholicism↗
PubMed2003

The N-terminus is unstructured, but not dynamically disordered, in the complex between HK022 Nun protein and lambda-phage BoxB RNA hairpin.

The Nun protein of lambdoid phage HK022 excludes lambda-phage superinfection by blocking expression of genes downstream from the lambda nut sequences. Heteronuclear NMR studies have been performed on a Nun peptide comprising residues 1-49 bound to the nutR BoxB RNA. The pattern of (13)C chemical shifts indicates that the arginine-rich motif of Nun forms an induced alpha-helix, consisting of residues 23-43, when bound to BoxB RNA, consistent with the structure of a shorter (residues 22-44) Nun peptide/BoxB RNA complex [Faber, C., Schärpf, M., Becker, T., Sticht, H. and Rösch (2001) J. Biol. Chem. 276, 32064-32070]. The N-terminal extension, residues 1-22, does not show chemical shifts or nuclear Overhauser effects characteristic of stable secondary structure. Nonetheless, (15)N relaxation measurements indicate that this region is not completely disordered, as expected for a random coil peptide. Restriction of conformation flexibility in the N-terminal extension of Nun may be important for binding to other target molecules involved in transcription termination.

Amino Acid Motifs↗
PubMed2001

The structure of the coliphage HK022 Nun protein-lambda-phage boxB RNA complex. Implications for the mechanism of transcription termination.

Nun protein from coliphage HK022 binds to phage boxB RNA and functions, in contrast to phage lambda N protein, as a transcriptional terminator. The basic Nun-(10-44) peptide contains the boxB RNA binding arginine rich motif, ARM. The peptide binds boxB RNA and competes with the phage lambda ARM peptide N-(1-36) as indicated by nuclear magnetic resonance (NMR) spectroscopy titrations. In two-dimensional nuclear Overhauser enhancement spectroscopy experiments boxB RNA in complex with Nun-(20-44) exhibits the same pattern of resonances as it does in complex with N peptides containing the ARM, and we could show that Nun-(20-44) forms a bent alpha-helix upon binding to the boxB RNA. The structure of the boxB RNA-bound Nun-(20-44) was determined on the basis of 191 intra- and 30 intermolecular distance restraints. Ser-24 is anchored to the lower RNA stem, and stacking of Tyr-39 and A7 is clearly experimentally indicated. Arg-28 shows numerous contacts to the RNA stem. Leu-22, Ile-30, Trp-33, Ile-37, and Leu-41 form a hydrophobic surface, which could be a recognition site for additional host factors such as NusG. Such a hydrophobic surface area is not present in N-(1-36) bound to boxB RNA.

Amino Acid Sequence↗
PubMed1997

The Nun protein of bacteriophage HK022 inhibits translocation of Escherichia coli RNA polymerase without abolishing its catalytic activities.

Bacteriophage HK022 Nun protein blocks transcription elongation by Escherichia coli RNA polymerase in vitro without dissociating the transcription complex. Nun is active on complexes located at any template site tested. Ultimately, only the 3'-OH terminal nucleotide of the nascent transcript in an arrested complex can turn over; it is removed by pyrophosphate and restored with NTPs. This suggests that Nun inhibits the translocation of RNA polymerase without abolishing its catalytic activities. Unlike spontaneously arrested complexes, Nun-arrested complexes cannot be reactivated by transcription factor GreB. The various complexes show distinct patterns of nucleotide incorporation and pyrophosphorolysis before or after treatment with Nun, suggesting that the configuration of RNAP, transcript, and template DNA is different in each complex.

Bacteriophage lambda↗
PubMedDate not supplied

Blood pressure in nuns in a secluded order: A 30-year follow-up.

The powerful effect of psychosocial and acculturating influences on population blood pressure trends seems to be confirmed, through longitudinal observations, in the nuns in a secluded order. After initial observations had been made on culture, body form, blood pressure, diet, and other variables in 144 nuns and 138 lay women, included as a control group, a 30-year follow-up study was undertaken. Most striking were opposite trends noted between the two groups in blood pressure trend. During the follow-up period, blood pressure remained remarkably stable among the nuns. None showed an increase in diastolic blood pressure over 90 mm Hg. By contrast, the control women showed the expected increase in blood pressure with age. This resulted in a gradually greater difference (delta>30/15 mm Hg) in systolic and diastolic blood pressure between the two groups, which was statistically significant. In addition, cardiovascular morbidity and mortality, expressed as the outcome of fatal and nonfatal events, were different in the two groups. They were significantly more common in the lay women than in the nuns. Comparisons between survival curves were statistically significant (p = 0.0043 for fatal events; p = 0.0056 for nonfatal events) between the two groups. In conclusion, it seems reasonable to attribute much of the difference in blood pressure and cardiovascular events, to the different burden in psychosocial factor and to the preserved peaceful lifestyle of the nuns.

Adult↗
PubMedDate not supplied

An exploratory study of health practices of American Catholic nuns.

American Catholic nuns represent a distinct segment of American women, yet little is known about their health practices. Assessment of health-related activities of these women was conducted by means of a self-report survey which was completed by 345 nuns in the midwestern United States. Although 53% had a complete physical examination in the last year, 11% had not had one for more than seven years, and 44% had not had a recent breast or pelvic examination. Thirty-seven percent did monthly breast self-examination (BSE), 37% examined themselves rarely, and 26% never performed BSE. Nuns performing BSE were more likely to have been examined by a health care provider in the past year (p less than .05). Nuns reporting a more positive health status said they got an adequate amount of sleep and participated in regular exercise, yet reported themselves to be overweight (p less than .05). Stress was reported by 74%, and an inverse relationship between perceived relaxation time and perceived stress level was noted (p less than .05). Overall, data reflected certain health educational and behavioral deficits. Because nuns comprise a chronologically older cohort than their secular counterparts, efforts that enable adoption of wellness practices and facilitate improved health care among these women seem to be warranted.

Aged↗
PubMed1985

Blood pressure changes over 20 years in nuns in a secluded order.

Blood pressure was recorded for 20 years in 138 nuns in a secluded order and in 126 lay women taken as controls. During the study period none of the nuns or control women smoked, took oral contraceptives or changed residence. Diet was unrestricted and comparable in the two groups. Subjects of both groups were consecutively enrolled without any selection parameters. During the 20 years of observation, systolic and diastolic blood pressure rose with age in the control women but remained almost unchanged in the nuns. The mean of the regression slope of systolic and diastolic blood pressure versus age, which was calculated for each subject, was 0.089 and 0.054 respectively in the nuns, as opposed to 2.171 and 0.742 in the lay women (both P < 0.001). Secluded nuns, who spend almost all their time in silence and prayer in the virtual absence of company apart from when attending religious celebrations, thus showed almost no change in blood pressure with age over a 20-year period. The results support the hypothesis that, at least in women, everyday stress may be a determinant of the progressive increase in blood pressure with ageing.

Adult↗
PubMed1999

[Nuns and nursing care in the middle of the 20th century].

The aim of this qualitative study was to investigate the connection between the life of nine nuns in the Augustian order, simultaneously working as nurses in the middle of the 20th century. To get an extensive idea about this period, the nuns, now 60 to 84 years old, were asked about their life at that time in narrative interviews. They had all worked between 1930 and 1960 as nurses. Through content analysis according to Mayring (1997) the authors generated the core category "We have always been there". This category shows that nuns constantly had to be present for patients, nurses, medication and the head of the order, that they were held responsible for all care and that they had to be available for the order and the hospital all day and night. The head of the order determined where they had to work. These irregular working hours and the fact that they frequently took over management positions prematurely led young nuns beyond their physical and psychological limits. Their religion should serve as "source of effort" for their work as nurses.

Aged↗
PubMed1999

Binding of transcription termination protein nun to nascent RNA and template DNA.

The amino-terminal arginine-rich motif of coliphage HK022 Nun binds phage lambda nascent transcript, whereas the carboxyl-terminal domain interacts with RNA polymerase (RNAP) and blocks transcription elongation. RNA binding is inhibited by zinc (Zn2+) and stimulated by Escherichia coli NusA. To study these interactions, the Nun carboxyl terminus was extended by a cysteine residue conjugated to a photochemical cross-linker. The carboxyl terminus contacted NusA and made Zn2+-dependent intramolecular contacts. When Nun was added to a paused transcription elongation complex, it cross-linked to the DNA template. Nun may arrest transcription by anchoring RNAP to DNA.

Amino Acid Sequence↗
PubMed1987

The superior sleep of healthy elderly nuns.

The nocturnal sleep structure of ten healthy elderly nuns was compared to that of ten healthy age-matched female controls. The nuns fell asleep more quickly and had less early morning awakening, as well as greater REM sleep time. These differences may reflect the more highly entrained life style of the nuns, including modest habitual sleep restriction (and thus accumulated sleep debt) of about one hour each night. The current findings, reviewed in relation to the sleep and aging literature, tend to support the concept that some of the effects of aging on sleep can be offset by attention to good "sleep hygiene," including careful entrainment of sleep schedule and modest habitual sleep restriction. However, age-related decrements in slow-wave sleep and sleep maintenance were not significantly better in the nuns.

Aged↗
PubMed1997

Blood pressure trend and psychosocial factors: the case of the nuns in a secluded order.

The powerful effect of psychosocial and acculturating influences on population blood pressure trends seems to be confirmed, through longitudinal observations in the nuns in a secluded order. After initial observation had been made on culture, body form, blood pressure, diet, and other variables in 144 nuns and 138 lay-women, included as a control group, a 30-year follow-up study was undertaken. Most striking were opposite trends noted between the two groups in blood pressure trend. During the follow-up period blood pressure remained remarkably stable among the nuns. None showed a rise in diastolic blood pressure to above 90 mmHg. By contrast, the control women showed the expected increase in blood pressure with age. This resulted in a gradually greater difference (A > 30/15 mmHg) in systolic and diastolic blood pressure between the two groups, which was statistically significant. It appears reasonable to attribute much of the difference in blood pressure to the different burden in psychosocial factor and to the preserved peaceful lifestyle of the nuns.

Adult↗
PubMed2006

A pertussis outbreak associated with social isolation among elderly nuns in a convent.

The pertussis incidence during an outbreak in a convent in The Netherlands in 1992 was higher among 75 retired (unvaccinated) nuns (60%) than among 24 staff members (8%) and was higher among 9 nuns with only a convent career (100%) than among 66 nuns who had a career outside of the convent (55%). The pertussis incidence increased with duration of social isolation but not with age.

Aged↗
PubMed1991

'Nuns, virgins, and spinsters'. Rigoni-Stern and cervical cancer revisited.

The view that nuns have a very low risk of cervical cancer is questioned. The historical evidence for this view is reviewed, from the beginning of the eighteenth century to the present. An estimate of the actual mortality rate from cervical cancer suggests that risk of death from this neoplasm among nuns is little different from that among the general female population. It is recommended that nuns should not be excluded from cervical cytology screening. When symptoms which might suggest cervical cancer arise in such women, full gynaecological assessment is necessary.

Adult↗