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L Pillus

Publications and source records attributed to L Pillus.

31 records · Page 2Linked to original sources
PubMed1996

Yeast SAS silencing genes and human genes associated with AML and HIV-1 Tat interactions are homologous with acetyltransferases.

Silencing is an epigenetic form of transcriptional regulation whereby genes are heritably, but not necessarily permanently, inactivated. We have identified the Saccharomyces cerevisiae genes SAS2 and SAS3 through a screen for enhancers of sir1 epigenetic silencing defects. SAS2, SAS3 and a Schizosaccharomyces pombe homologue are closely related to several human genes, including one associated with acute myeloid leukaemia arising from the recurrent translocation t(8;16)(p11;p13) and one implicated in HIV-1 Tat interactions. All of these genes encode proteins with an atypical zinc finger and well-conserved similarities to acetyltransferases. Sequence similarities and yeast mutant phenotypes suggest that SAS-like genes function in transcriptional regulation and cell-cycle exit and reveal novel connections between transcriptional silencing and human disease.

Acetyltransferases↗
PubMed1996

Activation of an MAP kinase cascade leads to Sir3p hyperphosphorylation and strengthens transcriptional silencing.

During cell division and growth, the nucleus and chromosomes are remodeled for DNA replication and cell type-specific transcriptional control. The yeast silencing protein Sir3p functions in both chromosome structure and in transcriptional regulation. Specifically, Sir3p is critical for the maintenance of telomere structure and for transcriptional repression at both the silent mating-type loci and telomeres. We demonstrate that Sir3p becomes hyperphosphorylated in response to mating pheromone, heat shock, and starvation. Cells exposed to pheromone arrest in G1 of the cell cycle, yet G1 arrest is neither necessary nor sufficient for pheromone-induced Sir3p hyperphosphorylation. Rather, hyperphosphorylation of Sir3p requires the mitogen-activated protein (MAP) kinase pathway genes STE11, STE7, FUS3/KSS1, and STE12, indicating that an intact signal transduction pathway is crucial for this Sir3p phosphorylation event. Constitutive activation of the pheromone-response MAP kinase cascade in an STE11-4 strain leads to hyperphosphorylation of Sir3p and increased Sir3p-dependent transcriptional silencing at telomeres. Regulated phosphorylation of Sir3p may thus be a mechanistically significant means for modulating silencing. Together, these observations suggest a novel role for MAP kinase signal transduction in coordinating chromatin structure and nuclear organization for transcriptional silencing.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1995

The SIR2 gene family, conserved from bacteria to humans, functions in silencing, cell cycle progression, and chromosome stability.

Genomic silencing is a fundamental mechanism of transcriptional regulation, yet little is known about conserved mechanisms of silencing. We report here the discovery of four Saccharomyces cerevisiae homologs of the SIR2 silencing gene (HSTs), as well as conservation of this gene family from bacteria to mammals. At least three HST genes can function in silencing; HST1 overexpression restores transcriptional silencing to a sir2 mutant and hst3 hst4 double mutants are defective in telomeric silencing. In addition, HST3 and HST4 together contribute to proper cell cycle progression, radiation resistance, and genomic stability, establishing new connections between silencing and these fundamental cellular processes.

Amino Acid Sequence↗
PubMed1993

SIR3 and SIR4 proteins are required for the positioning and integrity of yeast telomeres.

Heritable inactivation of genes occurs in specific chromosomal domains located at the silent mating type loci and at telomeres of S. cerevisiae. The SIR genes (for silent information regulators) are trans-acting factors required for this repression mechanism. We show here that the SIR3 and SIR4 gene products have a sub-nuclear localization similar to the telomere-associated RAP1 protein, which is found primarily in foci at the nuclear periphery of fixed yeast spheroplasts. In strains deficient for either SIR3 or SIR4, telomeres lose their perinuclear localization, as monitored by RAP1 immunofluorescence. The length of the telomeric repeat shortens in sir3 and sir4 mutant strains, and the mitotic stability of chromosome V is reduced. These data suggest that SIR3 and SIR4 are required for both the integrity and subnuclear localization of yeast telomeres, the loss of which correlates with loss of telomere-associated gene repression.

Cell Compartmentation↗
PubMed1992

An acquired state: epigenetic mechanisms in transcription.

Even when we know the primary sequences and binding specificities of every transcriptional activator and repressor, our understanding of transcriptional regulation will be rudimentary. This is partly because major aspects of gene expression are governed by epigenetic mechanisms. These mechanisms may be responsible for apparently identical sequences being read by the transcriptional machinery in two different but heritable ways: active or repressed. Epigenetic transcriptional states have been encountered in a number of recent experiments.

Animals↗
PubMed1989

Epigenetic inheritance of transcriptional states in S. cerevisiae.

SIR1, one of several genes required for repression of yeast silent mating type loci, has a unique role in repression of the HML alpha locus. Single-cell assays revealed that cells with mutant alleles of SIR1, including presumptive null alleles, existed as populations of genetically identical cells whose members were in one of two different regulatory states. A minority of cells had a repressed HML alpha locus whereas the majority had a derepressed HML alpha locus. The two states were mitotically stable, although rare changes in state were observed during mitotic growth, possibly reflecting heritable changes to the HML alpha locus at or before replication. Analysis of changes in state suggests that SIR1 protein has a role in the establishment but not the maintenance of repression of silent mating type genes, whereas SIR2, SIR3, and SIR4 are required for maintenance.

Alleles↗
PubMed1986

A chicken-yeast chimeric beta-tubulin protein is incorporated into mouse microtubules in vivo.

The role of divergent primary sequences in restricting tubulin function was tested in vivo by a gene transfection experiment. A chicken-yeast chimeric beta-tubulin DNA was introduced into 3T3 cells using the transfection vector pSV2. The 5' end of this gene, from chicken, is similar but not identical with that of mouse beta-tubulins; the 3' end, from yeast, contains a carboxyl terminus that is very different from other known beta-tubulin sequences. The chimeric protein is incorporated efficiently into each of the microtubule structures and each of the microtubules in the host cells. The presence of the protein has no apparent effect on either growth rate or cell morphology. The results suggest that the divergent sequences in this chimeric tubulin molecule place no restrictions on its activities in mouse cells.

Amino Acid Sequence↗
PubMed1986

Components of microtubular structures in Saccharomyces cerevisiae.

Most studies of cytoskeletal organelles have concentrated on molecular analyses of abundant and biochemically accessible structures. In many of the classical cases, however, the nature of the system chosen has precluded a concurrent genetic analysis. The mitotic spindle of the yeast Saccharomyces cerevisiae is one example of an organelle that can be studied by both classical and molecular genetics. We show here that this microtubule structure also can be examined biochemically. The spindle can be isolated by selective extractions of yeast cells by using adaptations of methods successfully applied to animal cells. In this way, microtubule-associated proteins of the yeast spindle are identified.

Benzimidazoles↗
PubMed1986

Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins.

Two alpha-tubulin genes from the budding yeast Saccharomyces cerevisiae were identified and cloned by cross-species DNA homology. Nucleotide sequencing studies revealed that the two genes, named TUB1 and TUB3, encoded gene products of 447 and 445 amino acids, respectively, that are highly homologous to alpha-tubulins from other species. Comparison of the sequences of the two genes revealed a 19% divergence between the nucleotide sequences and a 10% divergence between the amino acid sequences. Each gene had a single intervening sequence, located at an identical position in codon 9. Cell fractionation studies showed that both gene products were present in yeast microtubules. These two genes, along with the TUB2 beta-tubulin gene, probably encode the entire complement of tubulin in budding yeast cells.

Amino Acid Sequence↗
PubMed1983

Near-neighbor analysis of spindle microtubules in the alga Ochromonas.

The near-neighbor spacing of microtubules (MTs) in the spindle of the alga Ochromonas is analyzed. The technique of near-neighbor analysis of MTs (as developed by McDonald et al. [9]) in the mid-region of the Ochromonas spindle (overlap) shows that MTs from one pole preferentially associate with MTs from the opposite pole at a center-to-center distance of 35 to 43 nm. However, in the half spindle between the chromosomes and the poles, kinetochore MTs (kMTs) do not preferentially associate with other MTs in the half spindle but instead are arranged essentially at random. Individual polar MTs (MTs attached to one pole), kMTs and free MTs (MTs unattached to the poles) were selected for near-neighbor analysis over their entire lengths. The spacing of MTs in the overlap is compatible with those models for mitosis which propose that separation of the poles is accomplished by sliding between closely spaced MTs of opposite polarity. In contrast to the overlap, the arrangement of MTs in the half spindle is not compatible with MT2MT sliding theories that propose that chromosome movement is accomplished by sliding between kMTs and polar MTs.

Anaphase↗
PubMed1980

Organization of spindle microtubules in Ochromonas danica.

The entire framework of microtubules (MTs) in the mitotic apparatus of Ochromonas danica is reconstructed (except at the spindle poles) from transverse serial sections. Eleven spindles were sectioned and used for numerical data, but only four were reconstructed: a metaphase, an early anaphase, a late anaphase, and telophase. Four major classes of MTs are observed: (a) free MTs (MTs not attached to either pole); (b) interdigitated MTs (MTs attached to one pole which laterally associate with MTs from the opposite pole); (c) polar MTs (MTs attached to one pole); (d) kinetochore MTs (kMTs). Pole-to-pole MTs are rare and may be caused by tracking errors. During anaphase, the kMTs, free MTs, and polar MTs shorten until most disappear, while interdigitated MTs lengthen. In the four reconstructed spindles, the number of MTs decreases between early anaphase and telophase from 881 to 285, while their average length increases from 1.66 to 4.98 micron. The total length of all the MTs in the spindle (placed end to end) remains at 1.42 +/- 0.04 mm between these stages. At late anaphase and telophase the spindle is comprised mainly of groups of interdigitated MTs. Such MTs from opposite poles form a region of overlap in the middle of the spindle. During spindle elongation (separation of the poles), the length of the overlap region does not decrease. These results are compatible with theories that suggest that MTs directly provide the force that elongates the spindle, either by MT polymerization alone or by MT sliding with concomitant MT polymerization.

Anaphase↗