PubMed Health⌕ Search

Biomedical subjects

J W Tamkun

Publications and source records attributed to J W Tamkun.

At least 19 recordsLinked to original sources
PubMed1995

The role of brahma and related proteins in transcription and development.

The differential transcription of Drosophila homeotic genes is maintained by the Polycomb and trithorax groups of regulatory proteins, many of which are thought to modulate chromatin structure. During the past year, studies of a trithorax group member, brahma, and related yeast and human proteins have suggested that they are components of huge complexes that assist DNA-binding regulatory proteins to overcome the repressive effects of chromatin on transcription.

Animals↗
PubMed1995

The SWI-SNF complex: a chromatin remodeling machine?

The SWI-SNF complex plays a key role in the regulation of eukaryotic gene expression. Genetic studies in the yeast Saccharomyces cerevisiae suggest that one role for the complex is to antagonize chromatin-mediated repression of transcription. Recent biochemical studies indicate that S. cerevisiae and putative human SWI-SNF complexes use the energy of ATP hydrolysis to disrupt nucleosome structure.

Animals↗
PubMed1995

The Drosophila snr1 and brm proteins are related to yeast SWI/SNF proteins and are components of a large protein complex.

During most of Drosophila development the regulation of homeotic gene transcription is controlled by two groups of regulatory genes, the trithorax group of activators and the Polycomb group of repressors. brahma (brm), a member of the trithorax group, encodes a protein related to the yeast SWI2/SNF2 protein, a subunit of a protein complex that assists sequence-specific activator proteins by alleviating the repressive effects of chromatin. To learn more about the molecular mechanisms underlying the regulation of homeotic gene transcription, we have investigated whether a similar complex exists in flies. We identified the Drosophila snr1 gene, a potential homologue of the yeast SNF5 gene that encodes a subunit of the yeast SWI/SNF complex. The snr1 gene is essential and genetically interacts with brm and trithorax (trx), suggesting cooperation in regulating homeotic gene transcription. The spatial and temporal patterns of expression of snr1 are similar to those of brm. The snr1 and brm proteins are present in a large (> 2 x 10(6) Da) complex, and they co-immunoprecipitate from Drosophila extracts. These findings provide direct evidence for conservation of the SWI/SNF complex in higher eucaryotes and suggest that the Drosophila brm/snr1 complex plays an important role in maintaining homeotic gene transcription during development by counteracting the repressive effects of chromatin.

Amino Acid Sequence↗
PubMed1995

Developmental genetical analysis and molecular cloning of the abnormal oocyte gene of Drosophila melanogaster.

Studies of the abnormal oocyte (abo) gene of Drosophila melanogaster have previously been limited to the analysis of a single mutant allele, abnormal oocyte1 (abo1). The abo1 mutation causes a maternal-effect lethality that can be partially rescued zygotically by the abo+ allele and by increasing the dosage of specific regions of heterochromatin denoted ABO. This report describes the properties of abo2, a new P-element-induced allele that allowed us to reexamine the nature of maternal-effect defect. Comparisons of the phenotype of progeny of abo1/abo1 and abo1/abo2 females show that the preblastoderm lethality previously described as a component of the abo mutant maternal effect results from a recessive fertilization defect associated with the abo1 chromosome. We demonstrate here that the abo-induced maternal effect lethality occurs predominately late in embryogenesis after cuticle deposition but before hatching. The phenocritical period for zygotic rescue by heterochromatin coincides with this period of late embryogenesis. We have used the abo2 mutation to map and molecularly clone the gene. We show that the abo gene is located in the 32C cytogenetic interval and identify the putative abo transcript from mRNA isolated from adult females. Using germline transformation, we show that a 9-kb genomic fragment to which the transcript maps, partially fulfills requirement for maternal and zygotic abo+ function.

Animals↗
PubMed1994

Genetic analysis of the brahma gene of Drosophila melanogaster and polytene chromosome subdivisions 72AB.

The brahma gene is required for activation of the homeotic genes of the Antennapedia and bithorax complexes in Drosophila. We have isolated and characterized 21 mutations in brahma. We show that both maternal and zygotic functions of brahma are required during embryogenesis. In addition, the severe abnormalities caused by loss of maternal brahma expression show that the homeotic genes are not the only targets for brahma activation. The complex pattern of interallelic complementation for the 21 brahma alleles suggests that brahama may act as a multimer. In addition to mutations in brahma, we have isolated mutations in four other essential genes within polytene chromosome subdivisions 72AB. Based on a compilation of similar studies that include about 24% of the genome, we estimate that about 3600 genes in Drosophila can mutate to cause recessive lethality, with fewer than 900 additional genes essential only for gametogenesis. We have identified three times more transcripts than lethal complementation groups in 72AB. One transcript in 72AB is the product of the essential arf-like gene and encodes a member of the ARF subfamily of small GTP-binding proteins. Two other transcripts are probably the products of a single gene whose protein products are similar to the catalytic subunits of cAMP-dependent protein kinases.

ADP-Ribosylation Factors↗
PubMed1994

Identification and characterization of Drosophila relatives of the yeast transcriptional activator SNF2/SWI2.

The Drosophila brahma (brm) gene encodes an activator of homeotic genes that is highly related to the yeast transcriptional activator SWI2 (SNF2), a potential helicase. To determine whether brm is a functional homolog of SWI2 or merely a member of a family of SWI2-related genes, we searched for additional Drosophila genes related to SWI2 and examined their function in yeast cells. In addition to brm, we identified one other Drosophila relative of SWI2: the closely related ISWI gene. The 1,027-residue ISWI protein contains the DNA-dependent ATPase domain characteristic of the SWI2 protein family but lacks the three other domains common to brm and SWI2. In contrast, the ISWI protein is highly related (70% identical) to the human hSNF2L protein over its entire length, suggesting that they may be functional homologs. The DNA-dependent ATPase domains of brm and SWI2, but not ISWI, are functionally interchangeable; a chimeric SWI2-brm protein partially rescued the slow growth of swi2- cells and supported transcriptional activation mediated by the glucocorticoid receptor in vivo in yeast cells. These findings indicate that brm is the closest Drosophila relative of SWI2 and suggest that brm and SWI2 play similar roles in transcriptional activation.

Adenosine Triphosphatases↗
PubMed1993

BRG1 contains a conserved domain of the SWI2/SNF2 family necessary for normal mitotic growth and transcription.

Sequence-specific DNA binding activators of gene transcription may be assisted by SWI2 (SNF2), which contains a DNA-dependent ATPase domain. We have isolated a human complementary DNA encoding a 205K nuclear protein, BRG1, that contains extensive homology to SWI2 and Drosophila brahma. We report here that a SWI2/BRG1 chimera with the DNA-dependent ATPase domain replaced by corresponding human sequence restored normal mitotic growth and capacity for transcriptional activation to swi2- yeast cells. Point mutation of the conserved ATP binding site lysine abolished this complementation. This mutation in SWI2 exerted a dominant negative effect on transcription in yeast. A lysine to arginine substitution at the corresponding residue of BRG1 also generated a transcriptional dominant negative in human cells. BRG1 is exclusively nuclear and present in a high M(r) complex of about 2 x 10(6). These results show that the SWI2 family DNA-dependent ATPase domain has functional conservation between yeast and humans and suggest that a SWI/SNF protein complex is required for the activation of selective mammalian genes.

Adenosine Triphosphatases↗
PubMed1992

brahma: a regulator of Drosophila homeotic genes structurally related to the yeast transcriptional activator SNF2/SWI2.

The brahma (brm) gene is required for the activation of multiple homeotic genes in Drosophila. Loss-of-function brm mutations suppress mutations in Polycomb, a repressor of homeotic genes, and cause developmental defects similar to those arising from insufficient expression of the homeotic genes of the Antennapedia and Bithorax complexes. The brm gene encodes a 1638 residue protein that is similar to SNF2/SWI2, a protein involved in transcriptional activation in yeast, suggesting possible models for the role of brm in the transcriptional activation of homeotic genes. In addition, both brm and SNF2 contain a 77 amino acid motif that is found in other Drosophila, yeast, and human regulatory proteins and may be characteristic of a new family of regulatory proteins.

Adenosine Triphosphatases↗
PubMed1992

Trans-regulation of homeotic genes in Drosophila.

Homeotic genes of the Antennapedia and bithorax complexes control Drosophila development by encoding DNA-binding proteins that regulate the transcription of target genes. Because either the presence or absence of these DNA-binding proteins alters development, regulation of the spatial patterns of expression is crucial to normal development. Numerous gene products are required for properly regulated expression of Antennapedia and bithorax complex genes, but few (if any) are dedicated solely to the regulation of these genes. One of the pleiotropic activators of homeotic genes in Drosophila, the brahma gene, encodes a protein similar to a yeast protein that is required for transcriptional activation of multiple tightly regulated genes. Other components of this system may be conserved as well, suggesting that the biochemical basis for induced gene expression in single-celled organisms may have more in common with programmed developmental pathways in multicellular organisms than previously thought.

Animals↗
PubMed1991

The arflike gene encodes an essential GTP-binding protein in Drosophila.

We have identified a Drosophila gene (arflike, arl) encoding a protein that is structurally related (approximately 55% identity) to the ADP-ribosylation factors (ARFs) of yeast and mammals. Biochemical analyses of purified recombinant arl-encoded protein revealed properties similar to the ARF proteins, including the ability to bind and hydrolyze GTP. Clear functional differences between arl and ARF proteins, including a complete lack of ARF activity, suggest that arl is not a functional homolog of ARF. A recessive lethal arl mutation was recovered, demonstrating that the arl locus is an essential gene. We conclude that the arl locus encodes an essential member of the ARF subfamily of small GTP-binding proteins in Drosophila.

ADP-Ribosylation Factors↗
PubMed1988

Dosage-dependent modifiers of polycomb and antennapedia mutations in Drosophila.

Two genes known to control the determination of segmental identity in Drosophila melanogaster are polycomb and antennapedia. To identify additional genes involved in the determination of segmental identity, we have isolated dominant modifers (both suppressors and enhancers) of polycomb and/or antennapedia mutations. Sixty-four such modifier mutations have been recovered and mapped to 18 complementation groups. All of the mutations identify genes necessary for viability of the zygote. Six of the 18 genes that were identified by mutations that interact with polycomb and/or antennapedia have been previously characterized as homoeotic genes [i.e., Sex combs reduced (Scr), Brista (Ba), trithorax (trx), Polycomb (Pc), Polycomblike (Pcl), and Sex comb on midleg (Scm)]. Mutations in several of the additional loci identified here have also been shown to have homoeotic phenotypes.

Animals↗
PubMed1986

Cell-type-specific fibronectin subunits generated by alternative splicing.

Multiple fibronectin mRNAs arise by alternative splicing of the primary transcript of a single gene. We describe analyses of the contribution of this alternative splicing to fibronectin subunit heterogeneity in three different cell types using antisera directed against specific segments of fibronectin. beta-galactosidase-fibronectin fusion proteins produced with the lambda gt11 bacterial expression vector were used as immunogens. One region of alternative splicing accounts for differences in subunit size, while a second contributes to differences between the fibronectins present in blood plasma and in fibroblastic cells. We also show, however, that these two regions of alternative splicing do not account for all detectable subunits. We have also used these segment-specific antisera to show that blood platelets contain a spectrum of fibronectin subunits distinct from that found in blood plasma.

Animals↗
PubMed1986

Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin.

We describe the isolation, characterization, and sequence of cDNA clones encoding one subunit of the complex of membrane glycoproteins that forms part of the transmembrane connection between the extracellular matrix and the cytoskeleton. The cDNA sequence encodes a polypeptide of 89 kd that has features strongly suggesting the presence of a large N-terminal extracellular domain, a single transmembrane segment, and a small C-terminal cytoplasmic domain. The extracellular domain contains a threefold repeat of a novel 40 residue cysteine-rich segment, and the cytoplasmic domain contains a tyrosine residue that is a potential site for phosphorylation by tyrosine kinases. We propose the name integrin for this protein complex to denote its role as an integral membrane complex involved in the transmembrane association between the extracellular matrix and the cytoskeleton.

Actins↗
PubMed1985

Repeating modular structure of the fibronectin gene: relationship to protein structure and subunit variation.

Analysis of the exon-intron structure of the rat fibronectin gene shows that exons correspond precisely with repeating structural units in the protein and that alternative use of some exons produces fibronectin subunits that differ by the presence or absence of certain structural modules. Secondary structure predictions suggest that the repeating structure of the protein is further subdivided into smaller structural units and that these also correspond with exons in the gene.

Animals↗
PubMed1984

Fibronectin: a versatile gene for a versatile protein.

We have isolated cDNA and genomic clones for rat fibronectins. A single gene gives rise to three different mRNA species by alternative splicing at a complex intron-exon boundary within the coding region. The fibronectins encoded by these three mRNAs differ by the insertion of different protein segments in the C-terminal heparin-binding domain. The amino acid sequences of the cell-, heparin- and fibrin-binding domains in the C-terminal third of fibronectin were deduced from the DNA sequences. The cell- and heparin-binding regions each consist of several similar repeating sequences known as type III homologies, while the fibrin-binding region comprises three repeats of a different type (type I homologies). The sequences suggest several hypotheses for the structure-function relationships of these domains of fibronectin; these hypotheses are now being tested. Determination of intron-exon boundaries within the fibronectin gene is beginning to reveal the modular structure of the gene and its relation to the repeating structure of fibronectin.

Amino Acid Sequence↗