scholarly journals Primary sequence and developmental expression of a novel Drosophila melanogaster src gene.

1987 ◽  
Vol 7 (6) ◽  
pp. 2119-2127 ◽  
Author(s):  
R J Gregory ◽  
K L Kammermeyer ◽  
W S Vincent ◽  
S G Wadsworth
Keyword(s):  
Drosophila Melanogaster ◽  
Blot Hybridization ◽  
Structural Features ◽  
Developmental Expression ◽  
Reading Frame ◽  
Adult Females ◽  
Src Gene ◽  
Free Translation

We have sequenced a cDNA clone for the Drosophila melanogaster gene Dsrc28C, a homolog of the vertebrate gene c-src. The cDNA contains a single open reading frame encoding a protein of 66 kilodaltons which contains features highly conserved within the src family of tyrosine protein kinases. Novel structural features of the Dsrc28C protein include a basic pI and a polyglycine domain near the amino terminus. Cell-free translation of in vitro-transcribed RNA yielded a protein of the predicted size which could be immunoprecipitated by anti-v-src antisera. RNA blot hybridization revealed that the gene is expressed predominantly during embryogenesis, in imaginal disks of third-instar larvae, and in adult females. In situ hybridization showed that expression in adult females is largely confined to nurse cells and developing oocytes.

Primary sequence and developmental expression of a novel Drosophila melanogaster src gene

1987 ◽  
Vol 7 (6) ◽  
pp. 2119-2127
Author(s):  
R J Gregory ◽  
K L Kammermeyer ◽  
W S Vincent ◽  
S G Wadsworth
Keyword(s):  
Drosophila Melanogaster ◽  
Blot Hybridization ◽  
Structural Features ◽  
Developmental Expression ◽  
Reading Frame ◽  
Adult Females ◽  
Src Gene ◽  
Free Translation

We have sequenced a cDNA clone for the Drosophila melanogaster gene Dsrc28C, a homolog of the vertebrate gene c-src. The cDNA contains a single open reading frame encoding a protein of 66 kilodaltons which contains features highly conserved within the src family of tyrosine protein kinases. Novel structural features of the Dsrc28C protein include a basic pI and a polyglycine domain near the amino terminus. Cell-free translation of in vitro-transcribed RNA yielded a protein of the predicted size which could be immunoprecipitated by anti-v-src antisera. RNA blot hybridization revealed that the gene is expressed predominantly during embryogenesis, in imaginal disks of third-instar larvae, and in adult females. In situ hybridization showed that expression in adult females is largely confined to nurse cells and developing oocytes.

Delimiting regulatory sequences of the Drosophila melanogaster Ddc gene

1986 ◽  
Vol 6 (12) ◽  
pp. 4548-4557
Author(s):  
J Hirsh ◽  
B A Morgan ◽  
S B Scholnick
Keyword(s):  
Drosophila Melanogaster ◽  
Germ Line ◽  
Regulatory Elements ◽  
P Element ◽  
Developmental Expression ◽  
Upstream Region ◽  
Regulatory Sequences ◽  
Flanking Sequences

We delimited sequences necessary for in vivo expression of the Drosophila melanogaster dopa decarboxylase gene Ddc. The expression of in vitro-altered genes was assayed following germ line integration via P-element vectors. Sequences between -209 and -24 were necessary for normally regulated expression, although genes lacking these sequences could be expressed at 10 to 50% of wild-type levels at specific developmental times. These genes showed components of normal developmental expression, which suggests that they retain some regulatory elements. All Ddc genes lacking the normal immediate 5'-flanking sequences were grossly deficient in larval central nervous system expression. Thus, this upstream region must contain at least one element necessary for this expression. A mutated Ddc gene without a normal TATA boxlike sequence used the normal RNA start points, indicating that this sequences is not required for start point specificity.

Peroxisomal multifunctional enzyme type 2 from the fruitfly: dehydrogenase and hydratase act as separate entities, as revealed by structure and kinetics

2011 ◽  
Vol 435 (3) ◽  
pp. 771-781 ◽  
Author(s):  
Tatu J. K. Haataja ◽  
M. Kristian Koski ◽  
J. Kalervo Hiltunen ◽  
Tuomo Glumoff
Keyword(s):  
Crystal Structure ◽  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Structural Features ◽  
Full Length ◽  
Multifunctional Enzyme ◽  
Domain Composition ◽  
Substrate Channelling

All of the peroxisomal β-oxidation pathways characterized thus far house at least one MFE (multifunctional enzyme) catalysing two out of four reactions of the spiral. MFE type 2 proteins from various species display great variation in domain composition and predicted substrate preference. The gene CG3415 encodes for Drosophila melanogaster MFE-2 (DmMFE-2), complements the Saccharomyces cerevisiae MFE-2 deletion strain, and the recombinant protein displays both MFE-2 enzymatic activities in vitro. The resolved crystal structure is the first one for a full-length MFE-2 revealing the assembly of domains, and the data can also be transferred to structure–function studies for other MFE-2 proteins. The structure explains the necessity of dimerization. The lack of substrate channelling is proposed based on both the structural features, as well as by the fact that hydration and dehydrogenation activities of MFE-2, if produced as separate enzymes, are equally efficient in catalysis as the full-length MFE-2.

DNA from Drosophila melanogaster β-heterochromatin binds specifically to nuclear lamins in vitro and the nuclear envelope in situ

Gene ◽  
1996 ◽  
Vol 171 (2) ◽  
pp. 171-176 ◽  
Author(s):  
Ella A. Baricheva ◽  
Miguel Berrios ◽  
Sergei S. Bogachev ◽  
Igor V. Borisevich ◽  
Eugenia R. Lapik ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Nuclear Envelope ◽  
Nuclear Lamins

scholarly journals Expression of varicella-zoster virus VLT-ORF63 fusion transcript induces broad viral gene expression during reactivation from neuronal latency

2020 ◽  
Author(s):  
Werner J. D. Ouwendijk ◽  
Daniel P. Depledge ◽  
Labchan Rajbhandari ◽  
Tihana Lenac Rovis ◽  
Stipan Jonjic ◽  
...  
Keyword(s):  
Gene Expression ◽  
Varicella Zoster Virus ◽  
Viral Gene Expression ◽  
Fusion Transcript ◽  
Viral Gene ◽  
Reading Frame ◽  
Varicella Zoster ◽  
Rapid Amplification

SummaryVaricella-zoster virus (VZV) establishes lifelong neuronal latency in most humans world-wide, reactivating in one-third to cause herpes zoster and occasionally chronic pain. How VZV establishes, maintains and reactivates from latency is largely unknown. Latent VZV gene expression is restricted to VZV latency-associated transcript (VLT) and open reading frame 63 (ORF63) in naturally VZV-infected human trigeminal ganglia (TG). Notably, these transcript levels positively correlated suggesting co-regulated transcription during latency. Here, we used direct RNA-sequencing to identify fusion transcripts that combine VLT and ORF63 loci (VLT-ORF63) and are expressed during both lytic and latent VZV infections. Furthermore, real-time PCR, RNA in situ hybridization and 5’ rapid amplification of cDNA ends (RACE) all confirmed VLT-ORF63, but not canonical ORF63, expression in human TG. During lytic infection, one of the two major VLT-ORF63 isoforms encodes a novel fusion protein combining VLT and ORF63 proteins (pVLT-ORF63). In vitro, VLT is transcribed in latently VZV-infected human sensory neurons, whereas VLT-ORF63 expression is induced by reactivation stimuli. Moreover, the pVLT-ORF63-encoding VLT-ORF63 isoform induced transcription of lytic VZV genes. Collectively, our findings show that VZV expresses a unique set of VLT-ORF63 transcripts, potentially involved in the transition from latency to lytic VZV infection.

scholarly journals A cRNA Probe Detects Prunus Necrotic Ringspot Virus in Three Peach Cultivars after Micrografting and in Peach Shoots following Long-term Culture at 4 °C

HortScience ◽  
1999 ◽  
Vol 34 (2) ◽  
pp. 346-347 ◽  
Author(s):  
K. Heuss ◽  
Q. Liu ◽  
F.A. Hammerschlag ◽  
R.W. Hammond
Keyword(s):  
Prunus Persica ◽  
Blot Hybridization ◽  
Ringspot Virus ◽  
Shoot Cultures ◽  
Crna Probe ◽  
Reading Frame ◽  
Prunus Necrotic Ringspot Virus ◽  
Repeated Use ◽  
Graft Success

As part of a program to develop transgenic peach (Prunus persica L. Batsch) cultivars with resistance to Prunus necrotic ringspot virus (PNRSV), we are testing a system for measuring virus in peach shoot cultures. Micrografting in vitro is used for inoculation and slot-blot hybridization, with a digoxigenin (DIG)-labeled cRNA probe complementary to the 5′ open reading frame (ORF) of PNRSV RNA 3, for detection. In this study, we investigated whether infected shoots maintain virus infection over long periods of culture at 4 °C and if PNRSV-infected `Suncrest' shoot cultures can serve as graft bases to transmit virus equally well into cultivars Nemaguard, Springcrest, and Suncrest. The results of RNA hybridization analysis showed that virus was present in extracts of leaf samples from 2-year-old PNRSV-infected `Suncrest' shoots that had been subjected to varying lengths of incubation at 4 °C in the dark, suggesting that infected shoots can be maintained for repeated use. Rates of graft success were higher in heterografts between `Suncrest' bases and tips of `Springcrest' or `Nemaguard' than in autografts between `Suncrest' and `Suncrest', and there was equal efficacy of graft inoculation from `Suncrest' into these three cultivars.

scholarly journals State of populations of rare s pecies Silene jailensis N.I. Rubzov and Heracleum ligusticifolium M. Bieb. on Nikitskaya Yayla

2021 ◽  
Vol 1 (157) ◽  
pp. 114-122
Author(s):  
A. R. Nikiforov
Keyword(s):  
Adverse Effect ◽  
Small Area ◽  
Self Regulation ◽  
Structural Features ◽  
External Threats ◽  
Internal Mechanism ◽  
Viable Seeds ◽  
Destructive Processes

Populations of obligate petrophytes of the Mountain Crimea Heracleum ligusticifolium M. Bieb. (Apiaceae) and Silene jailensis N.I. Rubtzov (Caryophyllaceae) are distinguished by their small number due to the internal mechanism of their self-regulation: in small-area local stony habitats, plants of pregenerative age are regularly eliminated. This mechanis m operates independently of external threats, the adverse effect of which was smoothed out by the structural features of populations: the predominance of long -lived generative plants in S. jailensis, and the abundance of viable seeds in H. ligusticifolium. However, in recent years, the population of the Nikitskaya Yayla has been observed to have a weak lack of seed renewal of Silene jailensis and a complete lack of seed renewal of Heracleum ligusticifolium. This circumstance led to intra-population destructive processes. The probability of degradation and extinction of populations actualized the development of methods of reproduction and maintenance of plants of these species in vitro. By now, there is a stock of plants of these species that can be used for in situ reintroduction.

The casein kinase 1 alpha gene of Drosophila melanogaster is developmentally regulated and the kinase activity of the protein induced by DNA damage

1996 ◽  
Vol 109 (7) ◽  
pp. 1847-1856 ◽  
Author(s):  
J.A. Santos ◽  
E. Logarinho ◽  
C. Tapia ◽  
C.C. Allende ◽  
J.E. Allende ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Kinase Activity ◽  
Early Embryo ◽  
Casein Kinase ◽  
Early Embryogenesis ◽  
Kinase Gene ◽  
Casein Kinase 1 ◽  
Adult Females ◽  
Early Embryos

We report the molecular cloning and characterisation of the first CK1(casein kinase) gene of Drosophila melanogaster (dmCK1). The protein sequence (DMCK1) shares significant homology with other mammalian CK1 protein kinases of the alpha sub-class. The dmCK1 gene is expressed only in adult females and during early embryonic development as a single transcript. Western blot analysis of total protein extracts of different stages of development show that the gene product is likewise present during early embryogenesis and in adult females. Kinase activity studies show that DMCK1 is active when in vitro translated but inactive when immunoprecipitated from total early embryo extracts. However, after dephosphorylation treatment the immunoprecipitates show high kinase activity. More significantly, DMCK1 kinase activity present in the immunoprecipitates can be specifically activated by gamma-irradiation of early embryos. Also, when DMCK1 is immunoprecipitated after irradiation it appears to undergo phosphorylation. Immunolocalization of DMCK1 in early embryos shows that the protein is predominantly cytoplasmic but after irradiation there is a significant relocalization to the interphase nucleus. The results suggest a possible requirement of the Drosophila CK1 alpha for mechanisms associated with DNA repair during early embryogenesis.

scholarly journals Molecular cloning and mRNA expression analysis of two GH secretagogue receptor transcripts in orange-spotted grouper (Epinephelus coioides)

2008 ◽  
Vol 199 (2) ◽  
pp. 253-265 ◽  
Author(s):  
Ting Chen ◽  
Zhiguo Tang ◽  
Aifen Yan ◽  
Wensheng Li ◽  
Haoran Lin
Keyword(s):  
Amino Acids ◽  
Pituitary Gland ◽  
Feedback Regulation ◽  
Expression Level ◽  
Epinephelus Coioides ◽  
Reading Frame ◽  
Mrna Expression Analysis ◽  
Acyl Ghrelin

GH secretagogue receptor (GHSR) is the receptor of ghrelin, a circulating GH-releasing and appetite-inducing hormone. In this paper, two Ghsr cDNAs, gpGhsr1a and gpGhsr1b, were identified and characterized in a teleost, the orange-spotted grouper (Epinephelus coioides). The gpGHSR1a is 1512 bp in length with an open reading frame (ORF) that encodes a protein of 383 amino acids with seven transmembrane (TM) domains, while the 1703 bp gpGHSR1b contains an ORF encoding for 303 amino acids with five TM domains. Comparison between cDNA and gene sequences showed that the two transcripts are two alternative splicing forms of a single gpGhsr gene. Tissue distribution and ontogeny of two gpGhsr mRNAs were examined by RT-PCR. The gpGHSR1a is mainly expressed in brain and pituitary gland, when compared with a more widespread expression of gpGHSR1b. During embryonic and larval development, the gpGhsr1b mRNA appears before the gpGhsr1a mRNA. Furthermore, quantitative real-time PCR performed on brain showed that both transcripts have the highest expression level in the pituitary gland. The expression level of gpGHSR1a was generally higher than that of gpGHSR1b. GHSR expressing cells were also detected widely in grouper brain by in situ hybridization, with a broader distribution than previous reports in mammals. Finally, an in vitro study showed that expression of both gpGHSR transcripts in pituitary and hypothalamus is downregulated by GH and ghrelin but not by des-acyl ghrelin, and this suggests that feedback regulation of GHSR also exists in teleostean fishes.

scholarly journals IS1414, an Escherichia coliInsertion Sequence with a Heat-Stable Enterotoxin Gene Embedded in a Transposase-Like Gene

2000 ◽  
Vol 68 (10) ◽  
pp. 5710-5715 ◽  
Author(s):  
Annette McVeigh ◽  
Alessio Fasano ◽  
Daniel A. Scott ◽  
Sandra Jelacic ◽  
Steve L. Moseley ◽  
...  
Keyword(s):  
Burkholderia Cepacia ◽  
Rhizobium Meliloti ◽  
Structural Features ◽  
Protein Product ◽  
Toxin Gene ◽  
Gene Probe ◽  
Coding Region ◽  
Reading Frame ◽  
Heat Stable

ABSTRACT Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) was originally discovered in EAEC but has also been associated with enterotoxigenic E. coli (ETEC). Multiple genomic restriction fragments from each of three ETEC strains of human origin showed homology with an EAST1 gene probe. A single hybridizing fragment was detected on the plasmid of ETEC strain 27D that also encodes heat-stable enterotoxin Ib and colonization factor antigen I. We isolated and characterized this fragment, showing that it (i) carries an allele of astA nearly identical to that originally reported from EAEC 17-2 and (ii) expressed enterotoxic activity. Sequence analysis of the toxin coding region revealed thatastA is completely embedded within a 1,209-bp open reading frame (ORF1), whose coding sequence is on the same strand but in the −1 reading frame in reference to the toxin gene. In vitro expression of the predicted M r-∼46,000 protein product of ORF1 was demonstrated. ORF1 is highly similar to transposase genes of IS285 from Yersinia pestis, IS1356 from Burkholderia cepacia, and ISRm3 from Rhizobium meliloti. It is bounded by 30-bp imperfect inverted repeat sequences and flanked by 8-bp direct repeats. Based on these structural features, pathognomonic of a regular insertion sequence, this element was designated IS1414. Preliminary experiments to show IS1414 translocation were unsuccessful. Overlapping genes of the type suggested by the IS1414 core region have heretofore not been described in bacteria. It seems to offer a most efficient mechanism for intragenomic and horizontal dissemination of EAST1.

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