Biomedical subjects

S Hake

Publications and source records attributed to S Hake.

46 records · Page 3Linked to original sources

PubMed1993

EMAP, an echinoderm microtubule-associated protein found in microtubule-ribosome complexes.

The major non-tubulin polypeptide found associated with microtubules purified from unfertilized sea urchin eggs by cycles of pH-dependent assembly has a Mr of 77,000. The 77,000 Mr polypeptide is heat- and acid-labile, and is antigenically distinct from the mammalian brain MAPs, MAP-2 and tau. Affinity-purified antiserum against the 77,000 Mr polypeptide was used to survey a variety of cells and tissues for the presence of antigenically related polypeptides. A cross-reacting polypeptide, ranging in Mr from 72,000 to 80,000, was found in microtubule preparations from a wide variety of echinoderms, including sea urchins, starfish and sand dollars. Indirect immunofluorescence showed that the polypetide was found in interphase as well as mitotic microtubule arrays. No cross-reacting material was detected in microtubules isolated from marine molluscs, mammalian brain or mouse B16 cultured cells. Because the 77,000 Mr MAP is abundant in echinoderms, we have called it EMAP for echinoderm microtubule-associated protein. Although the precise function of the EMAP is not known, our data suggest that the EMAP is involved in the attachment of ribosomes to microtubules. Large numbers of ribosomes are attached to the walls of EMAP-containing microtubules, but not EMAP-deficient microtubules. Removal of the EMAP from the microtubule by salt-extraction results in the release of ribosomes from the microtubule, indicating that the EMAP may form part or all of the long tapered stalk that connects these two organelles.

Animals↗

PubMed1992

Unraveling the knots in plant development.

Homeobox genes, first discovered from studies of homeotic mutations in Drosophila, have recently been found in plants. The proteins encoded by homeobox genes thus join the ranks of other animal transcription factors that have plant developmental counterparts, suggesting that even though plant and animal development are very different, regulatory mechanisms that direct development may be shared among all higher eukaryotes. The role of homeobox genes in plants remains elusive; nonetheless, gain-of-function mutations of one homeobox gene, Knotted, profoundly affect development. The phenotype suggests that ectopic expression of Knotted in leaves causes cells to take on alternative fates.

Animals↗

PubMed1992

Active Mutator elements suppress the knotted phenotype and increase recombination at the Kn1-O tandem duplication.

The KNOTTED-1 (KN1) locus is defined by a number of dominant mutations that affect leaf development. The Kn1-O mutation is characterized by outpocketings of tissue along lateral veins of the maize leaf and by displacement of ligule tissue from the junction of the blade and sheath into the blade. Kn1-O results from a tandem duplication of 17 kb; each repeat includes the entire 8-kb KN1 transcription unit. Mutator (Mu) transposable elements inserted at the junction of the two repeats diminish the mutant phenotype. The Mu insertions affect the Kn1-O mutation in several distinctive ways. (1) Two of the three Mu elements, a Mu1 and a Mu8 element, diminish the mutant phenotype only when active as indicated by hypomethylation; when methylated or inactive, the phenotype is comparable to the Kn1-O progenitor. (2) Additional rearrangements have arisen in these derivatives that further reduce the mutant phenotype. (3) A 100-2000-fold increase in the loss of one repeat occurs in the presence of Mu elements as compared to Kn1-O without elements. The high frequency of loss only occurs when the Mu elements are hypomethylated. The frequency is also influenced by the specific allele carried at the same locus on the homologous chromosome. Reciprocal exchange of flanking markers does not accompany the loss events. Various recombination models that address the events occurring at Kn1-O are presented.

Alleles↗

PubMed1992

A dominant mutation in the maize homeobox gene, Knotted-1, causes its ectopic expression in leaf cells with altered fates.

Dominant mutations of the Knotted-1 (Kn1) homeobox gene of maize alter the differentiation and growth of cells associated with leaf veins. By analyzing Kn1 transcripts and KN1 protein, we show that the gene is not expressed at high levels during the development of wild-type leaves. Instead, Kn1 is expressed in apical meristems of vegetative and floral shoots, and is downregulated as leaves and floral organs are initiated. Kn1 is also expressed in relatively undifferentiated cells within developing vascular bundles, as well as ground tissue, in immature, unelongated axes of wild-type vegetative and floral shoots. In Kn1-N2 mutant plants, quantitative, but not qualitative differences are apparent in Kn1 transcripts and KN1 protein, consistent with previous observations that dominant Kn1 mutations map to non-coding regions of the gene. Kn1 is expressed ectopically in vascular bundles within developing mutant leaves in a pattern that correlates with the phenotypic alterations produced by the Kn1-N2 mutation. Thus, Kn1 apparently alters the fates of leaf cells in which it is ectopically expressed from an early stage of leaf development. Based on these observations, we hypothesize that Kn1 functions in its wild-type context as a regulator of cell determination.

Fluorescent Antibody Technique↗

PubMed1991

The developmental gene Knotted-1 is a member of a maize homeobox gene family.

The Knotted-1 (Kn1) locus is defined by several dominant gain-of-function mutations that alter leaf development. Foci of cells along the lateral veins do not differentiate properly, but continue to divide, forming outpocketings or knots. The ligule, a fringe normally found at the junction of leaf blade and sheath, is often displaced and perpendicular to its normal position. The phenotype is manifested in all cell layers of the leaf blade, but is controlled by a subgroup of cells of the inner layer. Mutations result from the insertion of transposable elements or a tandem duplication. We show that the Kn1 gene encodes a homeodomain-containing protein, the first identified in the plant kingdom. Sequence comparisons strongly suggest that Kn1 acts as a transcription factor. Here we use the Kn1 homeobox to isolate other expressed homeobox genes in maize. The Kn1 homeobox may permit the isolation of genes that, like animal and fungal counterparts, regulate cell fate determination.

Amino Acid Sequence↗

PubMed1990

Mutant characters of knotted maize leaves are determined in the innermost tissue layers.

Knotted (Kn1), a dominant mutation in maize, perturbs normal leaf development. Mutant leaves have localized regions of extra growth called knots and, in addition to the normal ligule, ectopic fringes of ligule are found on the leaf blade. Previous clonal analysis showed that the epidermal genotype was immaterial in knot formation. To establish which inner leaf layer was required for formation of knots and ectopic ligule we used a closely linked albino mutation to mark X-ray-induced clonal sectors of wild type (kn) tissue in Kn1 plants. The sectors examined frequently changed in composition of layers in the leaf both transversely and longitudinally. We present results that show that both mutant characters are determined in the middle mesophyll-bundle sheath (MMBS) layer. We show that a lateral vein can produce a knot when only half the MMBS layer around the lateral vein contains the mutant gene. We also show that the ectopic ligule in Kn1 has contributions from both the adaxial epidermal and adaxial mesophyll layer.

In Vitro Techniques↗

PubMed1990

A tandem duplication causes the Kn1-O allele of Knotted, a dominant morphological mutant of maize.

Molecular and genetic techniques are used to define Kn1-O, a mutation which interferes with the normal differentiation of vascular tissue in leaves. Sequences associated with a previously cloned allele, Kn1-2F11, were used as hybridization probes in a Southern analysis of Kn1-O. By this analysis, Kn1-O lacks the Ds2 transposable element that causes Kn1-2F11 but instead is associated with a sequence duplication. Sequence and restriction analysis of genomic clones show that the duplication consists of a tandem array of two 17-kb repeats. Analysis of Kn1-O derivatives indicates that the duplication itself conditions the mutant phenotype; a severely knotted line, Kn1-Ox, has gained a repeat unit to form a triplication, whereas normal derivatives have either lost a repeat unit or sustained insertions that disrupt the tandem duplication. These insertions map near the central junction of the tandem duplication, suggesting that the mutant phenotype results from the novel juxtaposition of sequences. We discuss models that relate the tandem duplication of sequences to altered gene expression.

Alleles↗

PubMed1989

Cloning Knotted, the dominant morphological mutant in maize using Ds2 as a transposon tag.

The Kn1-2F11 mutation causes protrusions or knots along the lateral veins of the first few leaves of the maize plant. The phenotype is visible when an unlinked gene, presumably Ac, is present in the genome. The mutation is closely linked to a genetically unstable Adh1 mutation that resulted from the insertion of a Ds2 element (Döring et al., 1984; Chen et al., 1986). Using a unique sequence from the Ds2 element as a hybridization probe, a genomic restriction fragment that cosegregated with the knotted phenotype was cloned. It carries the Kn1-2F11 locus by the following criteria. (i) Cosegregation of the fragment is tightly linked to the phenotype. (ii) Somatic and germinal excision produce a fragment which is the expected size of a revertant fragment; progeny containing the revertant size fragment are normal. (iii) The sequences that hybridize to this fragment are significantly altered in the chromosome containing the original knotted mutation, Kn1-O, (iv) The cloned fragment does not hybridize to a chromosome that contains a deletion of Kn1-O.

Journal Article↗

PubMed1989

Nucleotide sequence of a cDNA encoding rat protamine and the haploid expression of the gene during rat spermatogenesis.

The nucleotide sequence of a 342-base cDNA encoding the rat protamine has been determined. This insert, isolated from a rat testis cDNA library, encodes a polypeptide of 50 amino acids of which 29 are arginine 9 are cysteine and 2 are tyrosine. The insert contains the complete 3'-noncoding region of 170 bases and 18 bases of the 5'-noncoding region. Hybridization of the protamine cDNA with the RNA prepared from testes of prepubertal and sexually mature rats revealed that protamine mRNA is first detectable as a 600 nucleotide long molecule in the 35-day old testis containing around 15% of round spermatids but not in testis of younger animals. The RNA of 50-day old and sexually mature rats was found to contain a second protamine mRNA which is around 500 nucleotides in length. Hybridization of the protamine cDNA with the RNA of isolated spermatids of the mature testis resulted in 2 prominent hybridization signals (600 and 500 bp) while the faint signal obtained with the RNA of pachytene spermatocytes (600 bp) was found to be due to contamination of the cell preparation by spermatids. After digestion of the mRNAs with ribonuclease H a single hybridization band even smaller than 500 nucleotides was obtained. As demonstrated on testis sections the transcripts are confined to the central layers of the tubuli seminiferi corresponding to the spatial arrangement of corresponding to the spatial arrangement of postmeiotic cells. The results indicate that the protamine gene in the rat is postmeiotically expressed and that the mRNA undergoes post-transcriptional processing that includes a reduction in molecular size with respect to the poly-(A)+ tail.

Amino Acid Sequence↗

PubMed1985

Coordinate induction of alcohol dehydrogenase 1, aldolase, and other anaerobic RNAs in maize.

Anaerobiosis results in the selective synthesis of a particular set of polypeptides in the maize root including the two alcohol dehydrogenases (Sachs, M. M., Freeling, M., and Okimoto, R. (1980) Cell 20, 761-768), pyruvate decarboxylase (Wignarajah, K., and Greenway, H. (1976) New Phytol. 77, 575-584; Laszlo, A., and St. Lawrence, P. (1983) Mol. Gen. Genet. 192, 110-117), glucose phosphate isomerase (Kelley, P. M., and Freeling, M. (1984) J. Biol. Chem. 259, 673-677) and aldolase (Kelley, P. M., and Freeling, M. (1984) J. Biol. Chem. 259, 14180-14183). This report describes the identification and characterization of cDNA clones to five different mRNA species induced upon anaerobic shock. Immunoprecipitation of hybrid-selected translation polypeptides has determined the identity of the cDNA clone for fructose-1,6-diphosphate aldolase mRNA. Quantitative hybridization analysis of anaerobic mRNAs using the cDNA clones has shown that there is not a simultaneous accumulation of anaerobic mRNAs. Upon reintroduction of air, the anaerobic mRNAs disappear rapidly and at approximately the same rate. A translocation line that generates progeny that contain 1, 2, and 3 doses of the long arm of chromosome one (1L) allowed us to test for clustering of the anaerobic genes; two of the anaerobic genes tested do not reside with Adh 1 and Phi 1 on the long arm of chromosome 1.

Alcohol Dehydrogenase↗