Search PubMed⌕ Search

Biomedical subjects

D Shore

Publications and source records attributed to D Shore.

At least 19 recordsLinked to original sources
PubMed1992

A RAP1-interacting protein involved in transcriptional silencing and telomere length regulation.

The yeast RAP1 protein is a sequence-specific DNA-binding protein that functions as both a repressor and an activator of transcription. RAP1 is also involved in the regulation of telomere structure, where its binding sites are found within the terminal poly(C1-3A) sequences. Previous studies have indicated that the regulatory function of RAP1 is determined by the context of its binding site and, presumably, its interactions with other factors. Using the two-hybrid system, a genetic screen for the identification of protein-protein interactions, we have isolated a gene encoding a RAP1-interacting factor (RIF1). Strains carrying gene disruptions of RIF1 grow normally but are defective in transcriptional silencing and telomere length regulation, two phenotypes strikingly similar to those of silencing-defective rap1s mutants. Furthermore, hybrid proteins containing rap1s missense mutations are defective in an interaction with RIF1 in the two-hybrid system. Taken together, these data support the idea that the rap1s phenotypes are attributable to a failure to recruit RIF1 to silencers and telomeres and suggest that RIF1 is a cofactor or mediator for RAP1 in the establishment of a repressed chromatin state at these loci. By use of the two-hybrid system, we have isolated a mutation in RIF1 that partially restores the interaction with rap1s mutant proteins.

Amino Acid Sequence↗
PubMed1992

Dissection of a carboxy-terminal region of the yeast regulatory protein RAP1 with effects on both transcriptional activation and silencing.

RAP1 is an essential sequence-specific DNA-binding protein in Saccharomyces cerevisiae whose binding sites are found in a large number of promoters, where they function as upstream activation sites, and at the silencer elements of the HMR and HML mating-type loci, where they are important for repression. We have examined the involvement of specific regions of the RAP1 protein in both repression and activation of transcription by studying the properties of a series of hybrid proteins containing RAP1 sequences fused to the DNA-binding domain of the yeast protein GAL4 (amino acids 1 to 147). GAL4 DNA-binding domain/RAP1 hybrids containing only the carboxy-terminal third of the RAP1 protein (which lacks the RAP1 DNA-binding domain) function as transcriptional activators of a reporter gene containing upstream GAL4 binding sites. Expression of some hybrids from the strong ADH1 promoter on multicopy plasmids has a dominant negative effect on silencers, leading to either partial or complete derepression of normally silenced genes. The GAL4/RAP1 hybrids have different effects on wild-type and several mutated but functional silencers. Silencers lacking either an autonomously replicating sequence consensus element or the RAP1 binding site are strongly derepressed, whereas the wild-type silencer or a silencer containing a deletion of the binding site for another silencer-binding protein, ABF1, are only weakly affected by hybrid expression. By examining a series of GAL4 DNA-binding domain/RAP1 hybrids, we have mapped the transcriptional activation and derepression functions to specific parts of the RAP1 carboxy terminus.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcohol Dehydrogenase↗
PubMed1991

Separation of transcriptional activation and silencing functions of the RAP1-encoded repressor/activator protein 1: isolation of viable mutants affecting both silencing and telomere length.

The repressor/activator protein 1 (RAP1) binds to the upstream activating sites of many genes, the silencer elements flanking the unexpressed mating-type loci HMR and HML, and the poly(C1-3A) sequences at telomeres, suggesting that RAP1 might have three distinct regulatory functions. To determine the in vivo role of RAP1 in repression of the HMR silent locus, we developed a screen to isolate rap1 mutants specifically defective in silencing. Fifteen independent mutants defining four different rap1 alleles were isolated. These alleles are defective to different extents in repression of an HMR locus containing a mutated, but fully functional, silencer. All four alleles are missense mutations in only three codons within a small C-terminal region of the gene. These silencing-defective mutants have no apparent growth defects, indicating that expression of the large number of essential genes that have promoters containing RAP1-binding sites is normal. A transcriptional silencing function of RAP1 can therefore be genetically separated from its presumably essential activation functions. Surprisingly, three of the silencing-defective rap1 alleles have significantly longer telomeres, suggesting that the function of RAP1 in both transcriptional silencing and telomere-length regulation may be related. In addition, we have demonstrated that increased gene dosage of either SIR1 or SIR4, two other factors required for silencing, suppresses the silencing defect of the rap1 mutants. The properties of SIR4 dosage suppression suggest that SIR4 protein may interact directly with RAP1 at silencers.

Alleles↗
PubMed1991

RAP1 protein activates and silences transcription of mating-type genes in yeast.

RAP1 is a sequence-specific DNA-binding protein essential for cell growth. The occurrence of RAP1-binding sites in many promoter regions, the mating-type gene silencer elements, and telomeres suggests that RAP1 has multiple functions in the cell. To assess its role in transcription, temperature-sensitive mutations in RAP1 were generated. Analysis of rap1ts strains provides evidence that RAP1 functions in both transcriptional activation and silencing of mating-type genes. Several observations indicate that rap1ts strains are defective in the expression of MAT alpha, whose upstream activation sequence (UAS) contains a RAP1-binding site. At nonpermissive temperatures, decreases in MAT alpha steady-state transcript levels can be detected in MAT alpha rap1ts strains. Furthermore, these strains are deficient in alpha-pheromone production and simultaneously express at least two alpha-specific genes. These phenotypes can be reversed by replacing the RAP1-binding site at MAT alpha with a binding site for the GAL4 transcriptional activator. Certain rap1ts alleles have an opposite effect on the silent mating-type locus HMR, which becomes partially derepressed at nonpermissive temperatures.

Alleles↗
PubMed1991

RAP1 is required for BAS1/BAS2- and GCN4-dependent transcription of the yeast HIS4 gene.

The major in vitro binding activity to the Saccharomyces cerevisiae HIS4 promoter is due to the RAP1 protein. In the absence of GCN4, BAS1, and BAS2, the RAP1 protein binds to the HIS4 promoter in vivo but cannot efficiently stimulate HIS4 transcription. RAP1, which binds adjacently to BAS2 on the HIS4 promoter, is required for BAS1/BAS2-dependent activation of HIS4 basal-level transcription. In addition, the RAP1-binding site overlaps with the single high-affinity HIS4 GCN4-binding site. Even though RAP1 and GCN4 bind competitively in vitro, RAP1 is required in vivo for (i) the normal steady-state levels of GCN4-dependent HIS4 transcription under nonstarvation conditions and (ii) the rapid increase in GCN4-dependent steady-state HIS4 mRNA levels following amino acid starvation. The presence of the RAP1-binding site in the HIS4 promoter causes a dramatic increase in the micrococcal nuclease sensitivity of two adjacent regions within HIS4 chromatin: one region contains the high-affinity GCN4-binding site, and the other region contains the BAS1- and BAS2-binding sites. These results suggest that RAP1 functions at HIS4 by increasing the accessibility of GCN4, BAS1, and BAS2 to their respective binding sites when these sites are present within chromatin.

Base Sequence↗
PubMed1990

Involvement of the silencer and UAS binding protein RAP1 in regulation of telomere length.

The yeast protein RAP1, initially described as a transcriptional regulator, binds in vitro to sequences found in a number of seemingly unrelated genomic loci. These include the silencers at the transcriptionally repressed mating-type genes, the promoters of many genes important for cell growth, and the poly[(cytosine)1-3 adenine] [poly(C1-3A)] repeats of telomeres. Because RAP1 binds in vitro to the poly(C1-3A) repeats of telomeres, it has been suggested that RAP1 may be involved in telomere function in vivo. In order to test this hypothesis, the telomere tract lengths of yeast strains that contained conditionally lethal (ts) rap1 mutations were analyzed. Several rap1ts alleles reduced telomere length in a temperature-dependent manner. In addition, plasmids that contain small, synthetic telomeres with intact or mutant RAP1 binding sites were tested for their ability to function as substrates for poly(C1-3A) addition in vivo. Mutations in the RAP1 binding sites reduced the efficiency of the addition reaction.

Base Sequence↗
PubMed1990

Violent crime arrest rates of White House Case subjects and matched control subjects.

White House Case subjects typically are schizophrenic patients who have been psychotically preoccupied with a prominent political figure. The authors report the arrest rates for murder, aggravated assault, other assault, and robbery among 192 male White House Case subjects 20-59 years old who were discharged from civil commitments in the early 1970s. These rates were compared with those for the same crimes in a random matched sample of arrested subjects and with the general population arrest rates. When rates were covaried for prior violent crime arrests, White House Case subjects with prior arrests had a significantly higher rate of total posthospitalization violent crime arrests than the matched control sample.

Adult↗
PubMed1989

Fetal neural development and schizophrenia.

The conference on Fetal Neural Development and Schizophrenia which was held in Washington, DC, May 31-June 1, 1988, focused on factors of possible etiological significance in fetal development. Schizophrenia researchers joined experts in brain imaging, neuropathological, and neurochemical changes in brain development and investigators of potential genetic and neurobehavioral causes of psychosis. The combined evidence suggested dysfunction in frontal and parieto-occipital neocortex, basal ganglia, hippocampus, and amygdala. Dopamine transmission was implicated both in basal ganglia deficits and in widespread neocortical disturbances. Viral infection, or excessive stress, during the second trimester of pregnancy, as well as obstetrical complications, minor physical anomalies, and brain defects, correlated positively with incidence of adult schizophrenia. Autonomic nonresponding, birth complications, and ventricular enlargement were found to be closely related to negative symptom schizophrenia in high-risk populations. A dual factor model of schizophrenia was suggested, where genetic and environmental influences combine to produce psychosis.

Brain↗
PubMed1989

Murder and assault arrests of White House cases: clinical and demographic correlates of violence subsequent to civil commitment.

The authors studied arrest records and clinical data on 217 persons formerly hospitalized as "White House Cases" because they were psychotically preoccupied with prominent political figures. Prior arrest for violent crime was the variable most strongly associated with arrest for violent crime after hospital discharge. Male gender and a history of weapons possession were also correlated with future violence. For those with prior violent crime arrests, hospital incidents requiring seclusion were also associated with later violence. For those without prior arrests, subsequent violence was associated with threats, living outside Washington, and command hallucinations. For those previously arrested for nonviolent crimes, only persecutory delusions were associated with later violence.

Commitment of Persons with Psychiatric Disorders↗
PubMed1988

Attenuation of ultrasound in suspensions of bovine muscle myofibrils and myosin.

The attenuation of 1.5-7 MHz ultrasound was measured over the pH range 3-7 in 100 mM KCl suspensions of bovine M. semitendinosus myofibrils, precipitated myosin and the residue of myofibrils after partial extraction of myosin. In all fractions attenuation showed a similar dependence on pH over the range 3-7, with a broad, substantial maximum in the region of pH 4.5-pH 5.5 and similar mass attenuation coefficients (per g protein). At pH 7 and 7 MHz these were 3.49 +/- 0.20 cm2 g-1 in the myofibrils, 3.26 +/- 0.31 cm2 g-1 in the myofibrilar residue and 2.83 +/- 0.68 cm2 g-1 in the precipitated myosin. Measurements at 5.3 MHz of precipitated myosin over a wider pH range revealed an attenuation titration curve similar to that previously observed in homogenates of muscle and muscle myofibrils, with substantial peaks at about pH 5 and 11.5, and a shoulder perhaps indicating a small underlying peak at about pH 8-9. Myosin dissolved in 800 mM KCl gave attenuation levels that were typically 50% lower than precipitated myosin e.g. at pH 7 and 7 MHz: 2.83 +/- 0.68 cm2 g-1 in the precipitated form, 1.29 +/- 0.10 cm2 g-1 in solution. These results indicated that: (a) attenuation by myosin filaments contributed substantially to the total attenuation in suspensions of myofibrils and (b) the peak in the myofibrilar attenuation is caused, or substantially contributed to, by a process taking place in the myosin component.

Animals↗
PubMed1988

Experimental estimation of the viscous component of ultrasound attenuation in suspensions of bovine skeletal muscle myofibrils.

It has been suggested that viscous losses, caused by the motion of myofibrils relative to their environmental fluid, could be the major cause of ultrasound attenuation in muscle. This Paper presents theoretical and experimental estimates of the viscous component of attenuation in suspensions of myofibrils. Experimental estimates were made by monitoring the effects of varying the viscosity and density of the suspending fluid and ranged from 0.55 to 0.72 cm2 g-1 protein at 7 MHz, when expressed as mass attenuation coefficients. This represented 16-22% of the total attenuation measured in suspensions. Corresponding theoretical calculations were lower: 0.44 and 0.21 cm2 g-1 protein, respectively, for suspensions with the myofibrils aligned along and across the direction of sound propagation. It was concluded that most of the attenuation was caused by other absorption processes.

Animals↗
PubMed1988

Attenuation of ultrasound in homogenates of bovine skeletal muscle and other tissues.

The attenuation of ultrasound in homogenates of bovine skeletal muscle and suspensions of myofibrils was measured over the frequency range 1.5-7 MHz, and found to be proportional to protein concentration in both. In the homogenates it varied with frequency and temperature in a similar way to the attenuation in post rigor muscle tissue; myofibrils showed a higher frequency dependence. The attenuation in homogenates of bovine muscle, liver and kidney and in suspensions of myofibrils was measured over the pH range 3.5-13, and each showed a peak at about pH 11.5. This was thought to be due to a proton transfer process between NH3+ groups on the tissue proteins and OH- ions in the suspending fluid. A substantial peak at about pH 5 in the muscle and myofibril suspensions was not observed in homogenates of liver and kidney and was thought to be due to components of muscle that are absent from the other tissues. Myofibrils suspended in percoll solution of density 1.05 g cm-3, chosen to match approximately the density of the myofibrils, showed a slightly lower attenuation over the pH range 5-7, but a pH dependence similar to that of the myofibrils suspended in saline. The difference in the attenuations may be interpreted as the viscous component of the attenuation due to relative motion between the myofibril and its surrounding saline. The peak at pH 5 did not, however, appear to be due to the viscous loss mechanism peaking due to maximum shrinkage (and therefore maximum density) of the myofibril near this pH.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Violent crime arrests and paranoid schizophrenia: the White House case studies.

We have previously reported on typically paranoid schizophrenic patients who attempted to see the President or other prominent American political figures based on hallucinations or delusional beliefs. By obtaining arrest records on these White House Cases (WHCs), we were able to determine which individuals had murder or assault arrests before and/or after their WHC hospitalizations. During the 9-12 years following discharge, 31 of the 217 male WHCs (for whom adequate clinical records were available) had murder or assault arrests. Demographic characteristics such as prior violent crime arrest and male gender proved to be much better predictors of future violence than clinical symptom, history, or behavior items. Hospital incidents requiring seclusion and a history of weapons possession were both associated with later violence in WHCs with prior violent crime arrests, while certain clinical symptoms (e.g., persecutory delusions and command hallucinations) may be linked to future violence in WHCs without prior violent crime arrests. These data need replication in other patient samples.

Chronic Disease↗