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T C Kaufman

Publications and source records attributed to T C Kaufman.

At least 19 recordsLinked to original sources
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

Structural complexity and evolutionary conservation of the Drosophila homeotic gene proboscipedia.

Mutations of the homeotic gene proboscipedia (pb) of Drosophila cause striking transformations of the adult mouthparts, to legs or antennae. We report here an analysis of the gene structure of pb. Coding sequences across a 34 kb interval yield, by alternative splicing, four identified mRNA forms which differ immediately upstream of the homeobox. As a consequence, the homeodomain is expected to reside in four different contexts in the predicted protein isoforms. Mammalian homologs of pb, human HOX-2H and murine Hox-2.8, were identified based on the similarities of their homeodomains (95% identity) and several other conserved motifs. Examination of a collection of pb mutant alleles with antisera directed against the N-terminal region, the center or the C-terminal region of the protein showed that, surprisingly, several partial loss-of-function pb alleles appear to generate partially functional proteins truncated at their C-termini. This suggests that a significant portion of the protein contributes quantitatively to pb function, but is partially dispensable. Finally, evolutionary considerations suggest that pb may be one of several ancient genes which preceded the process yielding the modern homeotic gene complexes.

Amino Acid Sequence↗
PubMed1992

Ectopic expression of the Drosophila homeotic gene proboscipedia under Antennapedia P1 control causes dominant thoracic defects.

A deletion mutation in the Antennapedia Complex of Drosophila melanogaster, Df(3R)SCBXL2, induces both dominant and recessive loss-of-function phenotypes. The deletion is associated with diminished function of proboscipedia (pb), a homeotic gene required for mouthparts formation. Df(3R)SCBXL2 also has associated dominant thoracic defects related to diminished expression of the homeotic Antennapedia (Antp) gene copy on the homologous chromosome. This is shown to be a consequence of ectopic pb expression in the thorax. Newly juxtaposed Antp sequences provide the pb gene on the deletion bearing chromosome with a second promoter, Antp P1, in addition to its own. Ectopic pb protein expression occurs under Antp P1 control, by alternate splicing, and results in diminished accumulation of Antp protein in the imaginal disc cells where Antp P1 is normally expressed. The analysis of this mutant chromosome thus demonstrates that pb protein is capable of participating in the negative regulation of a more posteriorly expressed homeotic gene, as well as serving a homeotic "selector" function in the head.

Animals↗
PubMed1992

Homeotic genes have specific functional roles in the establishment of the Drosophila embryonic peripheral nervous system.

The Drosophila embryonic peripheral nervous system (PNS) contains segment-specific spatial patterns of sensory organs which derive from the ectoderm. Many studies have established that the homeotic genes of Drosophila control segment specific characteristics of the epidermis, and more recently these genes have also been shown to control gut morphogenesis through their expression in the visceral mesoderm (Tremml, G. and Bienz, M. (1989), EMBO J. 8, 2677-2685). We report here the roles of homeotic genes in establishing the spatial patterns of sensory organs in the embryonic PNS. The PNS was examined in embryos homozygous for mutations in the homeotic genes Sex combs reduced (Scr), Antennapedia (Antp), Ultrabithorax (Ubx), abdominal-A (abd-A) and Abdominal-B (Abd-B) with antibodies that label specific subsets of sensory organs. Our results suggest that the homeotic genes have specific roles in establishing the correct spatial patterns of sensory organs in their normal domains of expression. In addition, we also report the effects of ectopic expression of the homeotic genes labial (lab), Deformed (Dfd), Scr, Antp or Ubx on the normal development of sensory organs in the embryonic PNS. Interestingly, while previous studies have concluded that ectopic expression of the homeotic genes Dfd, Scr and Antp has no effect on the segmental identity of the abdominal segments, our results demonstrate that this is not true. We show that ectopic expression of these genes does result in the disruption of the developing PNS in the abdomen. Our results are suggestive of a role for the homeotic gene products in regulating genes which are necessary for generating sensory progenitor cells in the developing PNS.

Animals↗
PubMed1991

The doublesex locus of Drosophila melanogaster and its flanking regions: a cytogenetic analysis.

The region of the third chromosome (84D-F) of Drosophila melanogaster that contains the doublesex (dsx) locus has been cytogenetically analyzed. Twenty nine newly induced, and 42 preexisting rearrangements broken in dsx and the regions flanking dsx have been cytologically and genetically characterized. These studies established that the dsx locus is in salivary chromosome band 84E1-2. In addition, these observations provide strong evidence that the dsx locus functions only to regulate sexual differentiation and does not encode a vital function. To obtain new alleles at the dsx locus and to begin to analyze the genes flanking dsx, 59 lethal and visible mutations in a region encompassing dsx were induced. These mutations together with preexisting mutations in the region were deficiency mapped and placed into complementation groups. Among the mutations we isolated, four new mutations affecting sexual differentiation were identified. All proved to be alleles of dsx, suggesting that dsx is the only gene in this region involved in regulating sexual differentiation. All but one of the new dsx alleles have equivalent effects in males and females. The exception, dsxEFH55, strongly affects female sexual differentiation, but only weakly affects male sexual differentiation. The interactions of dsxEFH55 with mutations in other genes affecting sexual differentiation are described. These results are discussed in terms of the recent molecular findings that the dsx locus encodes sex-specific proteins that share in common their amino termini but have different carboxyl termini. The 72 mutations in this region that do not affect sexual differentiation identify 25 complementation groups. A translocation, T(2;3)Es that is associated with a lethal allele in one of these complementation groups is also broken at the engrailed (en) locus on the second chromosome and has a dominant phenotype that may be due to the expression of en in the anterior portion of the abdominal tergites where en is not normally expressed. The essential genes found in the 84D-F region are not evenly distributed throughout this region; most strikingly the 84D1-11 region appears to be devoid of essential genes. It is suggested that the lack of essential genes in this region is due to the region (1) containing genes with nonessential functions and (2) being duplicated, possibly both internally and elsewhere in the genome.

Alleles↗
PubMed1991

A functional and structural analysis of the Sex combs reduced locus of Drosophila melanogaster.

We have undertaken a developmental genetic analysis of the homeotic gene Sex combs reduced (Scr) of Drosophila melanogaster by examining embryonic and adult phenotypes of mutations affecting Scr gene function. Molecular mapping of Scr breakpoint lesions has defined a segment of greater than 70 kb of DNA necessary for proper Scr gene function. This region is split by the fushi tarazu (ftz) gene, with lesions affecting embryonic Scr function molecularly mapping to the region proximal (5') to ftz and those exhibiting polyphasic semilethality predominantly mapping distal (3') to ftz. Gain-of-function mutations are associated with genomic rearrangements and map throughout the Scr locus. Our analysis has revealed that the Scr locus encompasses genetic elements that are responsible for functions in both the embryonic and larval to adult periods of development. From these studies, we conclude that Scr is a complex genetic locus with an extensive regulatory region that directs functions required for normal head and thoracic development in both the embryo and the adult and that the regulation of Scr during these two periods is distinct.

Alleles↗
PubMed1991

The homeotic gene Sex combs reduced of Drosophila melanogaster is differentially regulated in the embryonic and imaginal stages of development.

The Sex combs reduced (Scr) locus is unique among the genes contained within the Antennapedia complex (ANT-C) of Drosophila melanogaster in that it directs functions that are required for both cephalic and thoracic development in the embryo and the adult. Antibodies raised against protein encoded by Scr were used to follow the distribution of this gene product in embryos and imaginal discs of third instar larvae. Analysis of Scr protein accumulation in embryos hemizygous for breakpoint lesions mapping throughout the locus has allowed us to determine that sequences required for establishment of the Scr embryonic pattern are contained within a region of DNA that overlaps with the identified upstream regulatory region of the segmentation gene fushi tarazu (ftz). Gain-of-function mutations in Scr result in the presence of ectopic sex comb teeth on the first tarsal segment of mesothoracic and metathoracic legs of adult males. Heterozygous combinations of gain-of-function alleles with a wild-type Scr gene exhibit no evidence of ectopic protein localization in the second and third thoracic segments of embryos. However, mesothoracic and metathoracic leg imaginal discs can be shown to accumulate ectopically expressed Scr protein, implying a differential regulation of the Scr gene during these two periods of development. Additionally, we have found that the spatial pattern of Scr gene expression in imaginal tissues involved in the development of the adult thorax is governed in part by synapsis of homologous chromosomes in this region of the ANT-C. However, those imaginal discs that arise anteriorly to the prothorax do not appear to be sensitive to this form of gene regulation. Finally, we have demonstrated that the extent of Scr expression is influenced by mutations at the Polycomb (Pc) locus but not by mutant alleles of the zeste (z) gene. Taken together, our data suggests that Scr gene expression is differentially regulated both temporally and spatially in a manner that is sensitive to the structure of the locus.

Animals↗
PubMed1991

Rescue and regulation of proboscipedia: a homeotic gene of the Antennapedia Complex.

The extraordinary positional conservation of the homeotic genes within the Antennapedia and the Bithorax Complexes (ANT-C and BX-C) in Drosophila melanogaster and the murine Hox and human HOX clusters of genes can be interpreted as a reflection of functional necessity. The homeotic gene proboscipedia (pb) resides within the ANT-C, and its sequence is related to that of Hox-1.5. We show that two independent pb minigene P-element insertion lines completely rescue the labial palp-to-first leg homeotic transformation caused by pb null mutations; thus, a homeotic gene of the ANT-C can properly carry out its homeotic function outside of the complex. Despite the complete rescue of the null, the minigene expresses pb protein in only a subset of pb's normal domains of expression. Therefore, the biological significance of the excluded expression pattern elements remains unclear except to note they appear unnecessary for specifying normal labial identity. Additionally, by using reporter gene constructs inserted into the Drosophila genome and by comparing pb-associated genomic sequences from two divergent species, we have located cis-acting regulatory elements required for pb expression in embryos and larvae.

Animals↗
PubMed1991

Developmental and evolutionary implications of labial, Deformed and engrailed expression in the Drosophila head.

Prior developmental genetic analyses have shown that labial (lab) and Deformed (Dfd) are homeotic genes that function in the development of the embryonic (larval) and adult head. Using antibody probes to reveal the spatial distribution of the lab and Dfd proteins in embryonic and imaginal tissues, we have assessed the respective roles of these genes through an analysis of the correspondence of their expression patterns with their mutant phenotypes. With regard to imaginal development, lab and Dfd occupy adjacent non-overlapping expression domains in the peripodial cell layer of the eye-antennal disc, in patterns that are consistent with their adult mutant phenotypes and published fate maps. During embryogenesis, lab and Dfd exhibit limited overlapping expression in areas that are of no obvious significance to the development of larval head structures, but also in areas that may have consequences for imaginal development. The head of Drosophila and other cyclorrhaphous Dipterans is characterized by an extreme morphological difference between the larval and adult stages. Given this unique ontogenetic and phylogenetic history and the observation that homeotic transformations produced by the lab, Dfd, and proboscipedia (pb) loci are manifested only in the adult, we suggest that distinct regulatory paradigms evolved for homeotic gene function in the development of the larval versus adult head. Finally, a detailed examination of the engrailed (en) expression pattern in the embryonic head strengthens the view of insect morphologists that the clypeolabrum evolved from the fusion of paired labral appendages.

Animals↗
PubMed1991

Control of expression of the homeotic labial (lab) locus of Drosophila melanogaster: evidence for both positive and negative autogenous regulation.

The homeotic gene labial (lab) is required for proper development of the embryonic and adult head in Drosophila melanogaster. The lab gene product accumulates in a complex pattern in both embryonic and imaginal tissue. During embryogenesis, lab is expressed in the endodermally derived cells of the midgut, in ectodermally derived cells of the procephalon and dorsal ridge, and in a small subset of progenitor sensory cells. Imaginal expression is restricted to a narrow region of the peripodial membrane of the eye-antennal disc. As part of our continuing effort to understand the role of lab in development, we have begun a dissection of the regulatory elements of the lab transcription unit and used germ line transformation experiments to determine which aspects of the observed expression pattern are essential for proper head development and viability. Transgenic embryos harboring an abridged lab gene are able to overcome the embryonic lethality associated with the loss of lab function and survive to adulthood. Interestingly, in these transgenic lines the lab protein accumulates only in a subset of those embryonic cells that normally express the gene, namely the procephalon and the anterior midgut. We also find that, once initiated, lab expression is maintained by positive autoregulation. Although lab minigene activity is sufficient to rescue the embryonic lethality of lab mutations, the transgenes fail to rescue defects in the adult head capsule. However, the defects observed in this study encompass a broader domain than those seen using somatic recombination to generate lab- clonal tissue. The failed rescue and observed cuticular defects are, at least in part, explained by the observation that the transgenes, rather than failing to be expressed, are associated with ectopic accumulation of lab protein in the peripodial membrane of the antennal disc. Moreover, this aberrant expression pattern is correlated with the abnormal expression of two other homeotic genes, Deformed (Dfd) and Sex combs reduced (Scr) in the eye-antennal disc. These results are only observed when the transgene is resident in a lab- genotype and ectopic expression of lab and misregulation of Dfd and Scr are not seen in a lab+ background. This result suggests that the wild-type lab gene product is necessary for the normal regulation of the locus in the imaginal discs, but unlike the case in the embryo, the event is negative. We discuss the biological implications of these results in relation to the role of lab in development.

Animals↗
PubMed1990

Molecular analysis of the bicoid gene from Drosophila pseudoobscura: identification of conserved domains within coding and noncoding regions of the bicoid mRNA.

The specification of anterior positional information during Drosophila embryogenesis is largely dependent upon the function of the maternal-effect gene bicoid (bcd). Two aspects of bcd function are particularly striking. First, the bcd protein product forms a gradient during early embryogenesis, which regulates the transcription of at least one zygotic segmentation gene, hunchback, in a concentration dependent manner. Secondly, formation of the bcd protein gradient is dependent upon the specific localization of bcd mRNAs at the anterior end of the oocyte/embryo during oogenesis, a process which requires a cis-acting 625 nucleotide sequence within the 3' untranslated region of the bcd mRNA. We have cloned and sequenced the bcd gene from Drosophila pseudoobscura as a tool in identifying important functional domains within this transcription unit. DNA sequence comparisons reveal: (i) varying degrees of amino acid sequence conservation among the proposed functional domains of the bcd protein, (ii) the conservation of potential RNA secondary structures within the bcd mRNA localization element, and (iii) the maintenance of a short open reading frame within the 5' untranslated leader that may play a role in translational regulation. Finally, the D.pseudoobscura bcd gene partially rescues the phenotype of a bcd- mutation when placed into the D.melanogaster genome by germline transformation. The lack of full phenotypic rescue can be explained in part by the observed improper localization of the D.pseudoobscura bcd mRNA when expressed in D.melanogaster.

Amino Acid Sequence↗
PubMed1990

Functional implications of the unusual spatial distribution of a minor alpha-tubulin isotype in Drosophila: a common thread among chordotonal ligaments, developing muscle, and testis cyst cells.

Three of the four alpha-tubulin genes in Drosophila melanogaster are temporally regulated. mRNA from one of these genes, alpha 85E-tubulin, first appears in 6- to 8-hr embryos and persists, with marked fluctuations, through the end of pupal development. In adults, alpha 85E mRNA has been unequivocally identified only in testes. In the present study, isotype-specific antibodies have been used to localize alpha 85E tubulin protein in whole tissues. The results demonstrate a spatially restricted expression pattern of the alpha 85E gene that includes tissues of both ectodermal and mesodermal origins. Specifically, embryonic accumulation of alpha 85E tubulin is limited to support cells of chordotonal organs and the developing musculature of the viscera and body wall. In late third instar larvae, chordotonal organs and a subset of larval nerves, but not muscle, stain with anti-alpha 85E. The timing of protein accumulation during pupal development suggests that alpha 85E tubulin is involved in the construction of the adult as well as the larval musculature. In testis, only the somatically derived cyst cells that surround developing spermatid bundles accumulate alpha 85E-tubulin. The cell types that express alpha 85E share a requirement for extensive cell shape changes during development, suggesting that this minor alpha-tubulin may have distinct functional properties.

Animals↗
PubMed1989

A genetic and developmental analysis of mutations in labial, a gene necessary for proper head formation in Drosophila melanogaster.

We have undertaken a developmental genetic analysis of labial (lab), the most proximal gene in the Antennapedia complex (ANT-C) of Drosophila melanogaster. The terminal phenotype of mutant embryos was examined in cuticle preparations, in thin sections, and by scanning electron microscopy. These preparations revealed a failure of head involution and the loss or disruption of several head structures, including the salivary glands and the H-piece and ventral arm of the cephalopharyngeal apparatus. Although these structures are presumed to derive from the gnathocephalic segments, we argue that the observed defects are likely to be a secondary consequence of a failure of head involution. A function for lab in the development of the adult head was inferred from the phenotype of animals bearing hypomorphic alleles and from clones of lab- tissue generated by mitotic recombination. Two aspects of the mutant phenotype were manifested. Ventrally, a deletion and/or disruption of tissue occurred in the maxillary palp and vibrissae regions. Dorsally, the posterior head appeared to be transformed to a thoracic-like identity. Mutations in lab, like those in the Deformed and proboscipedia loci of the ANT-C, reveal a homoeotic phenotype only in the adult stage of the life cycle.

Animals↗
PubMed1989

Developmental distribution of RNA and protein products of the Drosophila alpha-tubulin gene family.

The developmental pattern of gene expression of the Drosophila melanogaster alpha-tubulin family has been examined in detail at both the mRNA and protein levels. Northern data from 16 stages of development have been quantified to produce estimates of pool sizes of each of the alpha-tubulin transcripts through development. The in situ pattern of alpha 67C RNA localization in developing oocytes and early embryos has also been determined. At the protein level, two of the three previously unidentified products of alpha-tubulin genes (alpha 67C and alpha 85E) have been identified. Evidence that protein from the fourth gene comigrates with the ubiquitously expressed alpha 84B is presented. In addition to the primary translational products of the alpha-tubulin genes, an elaborate series of post-translationally modified alpha-tubulins has been resolved. The developmental profiles of both synthesis and accumulation of these alpha-tubulin proteins are described.

Animals↗
PubMed1989

Isolation, structure, and expression of labial, a homeotic gene of the Antennapedia Complex involved in Drosophila head development.

The labial (lab) gene of Drosophila melanogaster is necessary for the proper development of the embryonic (larval) and adult head. We have identified the lab transcription unit within the proximal portion of the Antennapedia Complex (ANT-C) by mapping the molecular lesions associated with chromosomally rearranged lab alleles. We present its molecular structure, nucleotide sequence, and temporal pattern of expression. In addition, using antibodies generated against a fusion protein, we show that in the embryo the lab protein is distributed in neural and epidermal cells of the procephalic lobe; in a discrete loop of the midgut; and in specific progenitor sensory cells of the clypeolabrum, thoracic segments, and tail region. The regions of lab expression in the developing cephalon represent nonsegmented domains that are anterior to and largely nonoverlapping with the domains of expression of the Deformed (Dfd) and proboscipedia (pb) genes, two other homeotic loci of the ANT-C that also function to direct the development of head structures. Furthermore, lab head expression is associated with the complex cellular movements of head involution, a process that not only is defective in lab embryos, but the failure of which appears to be largely responsible for the defects observed in mutant embryos. Finally, we suggest that lab head expression provides a molecular marker for an intercalary segment, an ancestral segment that has become morphologically indistinct during the evolution of the insect head.

Amino Acid Sequence↗
PubMed1989

Novel patterns of homeotic protein accumulation in the head of the Drosophila embryo.

Antibodies that specifically recognize proteins encoded by the homeotic genes: Sex combs reduced, Deformed, labial and proboscipedia, were used to follow the distribution of these gene products during embryogenesis. The position of engrailed-expressing cells was used as a reference and staining conditions were established that could distinguish, among cells expressing engrailed, one of the homeotic proteins or both. Our observations demonstrate two important facts about establishing identity in the head segments. First, in contrast to the overlapping pattern of homeotic gene expression in the trunk segments, we observe a non-overlapping pattern in the head for those homeotic proteins required during embryogenesis. In contrast, the spatial accumulation of the protein product of the non-vital proboscipedia locus overlaps partially with the distribution of the Deformed and Sex combs reduced proteins in the maxillary and labial segments, respectively. Second, two of the proteins, Sex combs reduced and Deformed, have different dorsal and ventral patterns of accumulation. Dorsally, these proteins are expressed in segmental domains while, within the ventral region, a parasegmental register is observed. The boundary where this change in pattern occurs coincides with the junction between the ventral neurogenic region and the dorsal epidermis. After contraction of the germ band, when the nerve cord has completely separated from the epidermis, the parasegmental pattern is observed only within the ventral nerve cord while a segmental register is maintained throughout the epidermis.

Animals↗