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R Lecanidou

Publications and source records attributed to R Lecanidou.

At least 19 recordsLinked to original sources
PubMed2005

BmCbZ, an insect-specific factor featuring a composite DNA-binding domain, interacts with BmC/EBPgamma.

A novel factor featuring a composite AT hook/basic region-leucine zipper DNA-binding domain was isolated from Bombyx mori follicular cells. Screening of EST databases derived from a variety of metazoans revealed the exclusive presence of BmCbZ homologues in insect species. BmCbZ characteristic features and gene organization are discussed, in comparison to other known bZIP factors. We concordantly propose that this factor establishes a new insect-specific bZIP family. We further present the isolation of the silkmoth homologue of mammalian C/EBPgamma, BmC/EBPgamma, and in vitro evidence for its interaction with BmCbZ. The formation of a BmCbZ-BmC/EBPgamma heterodimer is a prerequisite for binding to specific C/EBP recognition sites on chorion gene promoters, most probably via both major and minor groove interactions.

AT-Hook Motifs↗
PubMed2005

A novel role for the Bombyx Slbo homologue, BmC/EBP, in insect choriogenesis.

One previously unidentified cDNA clone coding for a C/EBP factor, BmC/EBP, was isolated from Bombyx mori follicular cells. This is the first time that a C/EBP factor has been isolated and characterized in Lepidoptera. We provide information concerning structural features and developmental specificity, as well as in vitro interaction properties with chorion gene promoter modules. BmC/EBP was capable of effectively recognizing homologous binding sites from chorion gene promoters derived from flies and other moths, despite significant diversity of chorion structure, gene organization, and gene expression profiles. We propose that the relative concentration of BmC/EBP, in relation to its differential binding affinity for promoter cis-elements, results in activation or repression of silkmoth chorion gene expression.

Animals↗
PubMed2003

In vitro analysis of Bombyx mori early chorion gene regulation: stage specific expression involves interactions with C/EBP-like and GATA factors.

This is the first attempt to identify regulatory elements that are involved in early choriogenesis of the silkworm Bombyx mori. A new cis element in the promoter region of five early chorion genes was identified. The consensus sequence of this element matches the consensus of the C/EBP DNA binding site. Moreover, this sequence interacts with a 70 kD protein (pX2) present in follicular nuclear extracts and complex formation exhibits early developmental specificity. There is strong evidence that this factor belongs to the C/EBP family. Surprisingly, the same protein binds with the same developmental specificity to a similar sequence of a late chorion gene promoter, which has been previously defined as the binding site for a putative late specific factor, BCFII. The possibility that pX2 and BCFII are isoforms or modifications of the same protein factor, which is presumably able to bind to the highly similar sequence elements of both early and late genes, is discussed. A hypothesis involving protein-protein interactions between C/EBP (pX2/BCFII) and GATA during choriogenesis is presented to explain the temporal specificity of chorion genes.

Animals↗
PubMed2001

The biolistic method as a tool for testing the differential activity of putative silkmoth chorion gene promoters.

Bombyx mori unpaired early chorion gene copies 6F6.1,.2 and.3 are exceptions to the typical organization and distribution pattern of known early ErA/ErB, middle A/B and late HcA/HcB divergently transcribed gene pairs. Contrary to such pairs, the boundaries of the 6F6 regulatory sequences are not easily defined; moreover, they share common sequence elements with the regulatory sequences of middle and late genes. In order to perform a functional study of the tissue and temporal specificity of the 6F6 putative promoter region, we decided to apply biolistics. In the present work, use of a region from the 6F6.2 5' untranslated sequence, spanning nucleotides -138 to the cap site, gave an expected expression pattern of a lacZ reporter gene. Temporal specificity was further verified by control experiments using the cloned intergenic sequence of the late gene pair HcA/B.12, which resulted in lacZ expression in late choriogenic follicles. At present, despite the recent successful germinal transgenesis of Bombyx mori, the biolistic transient expression system seems to be the most rapid technique to pursue the functional study of the promoter region of early chorion genes, including the three unconventional early 6F6 genes.

Animals↗
PubMed1995

Sequence analysis of a small early chorion gene subfamily interspersed within the late gene locus in Bombyx mori.

A comprehensive sequence analysis of three early chorion genes (6F6.1, 6F6.2, 6F6.3) which form a small subfamily is presented. Two main features characterize this subfamily: (1) the 6F6 gene copies are beta-branch genes and, unlike typical chorion genes which are organized in divergent gene pairs, they are unpaired, and (2) they are not clustered in genetic locus Ch3 but are dispersed in Ch1-2, which is about 3 to 4 centiMorgans away and contains middle and late chorion genes. Sequence comparisons show that members of this subfamily exhibit high identity values in their major coding region (94-96%) and that similarities also extend, but to a lesser degree, into their noncoding regions. The putative 6F6 promoter regions have no significant similarities with the corresponding regions of other early beta-genes but quite surprisingly share common elements with middle and late genes. The main difference among the 6F6 gene introns is the presence of inserted sequences: the insert into 6F6.2 ("IR"; 248 bp) is flanked by a 102-103-bp inverted repeat, while those into 6F6.1 ("FIB"; 184 bp) and 6F6.3 ("HOPE"; 951 bp) are carried by a partial Bm1 element. HOPE has features of a non-LTR retrotransposable element. Preliminary experiments indicate that the copy number of IR and HOPE in the Bombyx mori genome is about 5,000 and 20,000, respectively. The great similarity of 6F6 genes cannot be accounted for by selective pressure but rather appears to be the result of gene-conversion-like events, which are supposed to operate frequently in middle and late chorion genes but not in other known early beta-genes. Using the relative position and orientation of the 6F6 gene copies, it is possible to propose an evolutionary scheme for the formation of chorion locus Ch1-2.

Animals↗
PubMed1995

Complete sequence and gene organization of the mitochondrial genome of the land snail Albinaria coerulea.

The complete sequence (14,130 bp) of the mitochondrial DNA (mtDNA) of the land snail Albinaria coerulea was determined. It contains 13 protein, two rRNA and 22 tRNA genes. Twenty-four of these genes are encoded by one and 13 genes by the other strand. The gene arrangement shares almost no similarities with that of two other molluscs for which the complete gene content and arrangement are known, the bivalve Mytilus edulis and the chiton Katharina tunicata; the protein and rRNA gene order is similar to that of another terrestrial gastropod, Cepaea nemoralis. Unusual features include the following: (1) the absence of lengthy noncoding regions (there are only 141 intergenic nucleotides interspersed at different gene borders, the longest intergenic sequence being 42 nucleotides) (2) the presence of several overlapping genes (mostly tRNAs), (3) the presence of tRNA-like structures and other stem and loop structures within genes. An RNA editing system acting on tRNAs must necessarily be invoked for posttranscriptional extension of the overlapping tRNAs. Due to these features, and also because of the small size of its genes (e.g., it contains the smallest rRNA genes among the known coelomates), it is one of the most compact mitochondrial genomes known to date.

Animals↗
PubMed1994

Novel features of metazoan mtDNA revealed from sequence analysis of three mitochondrial DNA segments of the land snail Albinaria turrita (Gastropoda: Clausiliidae).

The mitochondrial DNA (mtDNA) size of the terrestrial gastropod Albinaria turrita was determined by restriction enzyme mapping and found to be approximately 14.5 kb. Its partial gene content and organization were examined by sequencing three cloned segments representing about one-fourth of the mtDNA molecule. Complete sequences of cytochrome c oxidase subunit II (COII), and ATPase subunit 8 (ATPase8), as well as partial sequences of cytochrome c oxidase subunit I (COI), NADH dehydrogenase subunit 6 (ND6), and the large ribosomal RNA (lrRNA) genes were determined. Nine putative tRNA genes were also identified by their ability to conform to typical mitochondrial tRNA secondary structures. An 82-nt sequence resembles a noncoding region of the bivalve Mytilus edulis, even though it might contain a tenth tRNA gene with an unusual 5-nt overlap with another tRNA gene. The genetic code of Albinaria turrita appears to be the same as that of Drosophila and Mytilus edulis. The structures of COI and COII are conservative, but those of ATPase8 and ND6 are diversified. The sequenced portion of the lrRNA gene (1,079 nt) is characterized by conspicuous deletions in the 5' and 3' ends; this gene represents the smallest coelomate lrRNA gene so far known. Sequence comparisons of the identified genes indicate that there is greater difference between Albinaria and Mytilus than between Albinaria and Drosophila. An evolutionary analysis, based on COII sequences, suggests a possible nonmonophyletic origin of molluskan mtDNA. This is supported also by the absence of the ATPase8 gene in the mtDNA of Mytilus and nematodes, while this gene is present in the mtDNA of Albinaria and Cepaea nemoralis and in all other known coelomate metazoan mtDNAs.

Amino Acid Sequence↗
PubMed1992

Three copies of the early gene 6F6 are interspersed in and around the late chorion gene cluster of Bombyx mori.

The developmentally regulated chorion genes of the silkmoth, Bombyx mori, are clustered in two genetic loci (Ch1-2 and Ch3) of chromosome 2, which are separated by approximately 4 centiMorgans. Early genes are clustered in chorion locus Ch3, whereas Ch1-2 contains all of the late genes in a continuous region of 140 kb, which is sandwiched between two regions containing most of the middle chorion genes. The late-gene area has been extensively studied and was considered to contain only late genes coding for chorion proteins of exceptionally high cysteine content organized in tightly clustered alpha/beta gene pairs. In the present paper we report the unexpected presence of three dispersed copies (6F6.1, 6F6.2, 6F6.3) of an early beta-gene, which disrupts the continuity of the late locus. Hybridization data indicate that 6F6.2 corresponds to the previously characterized m6F6 cDNA clone and that 6F6.1 and 6F6.3, but not 6F6.2, are adjacent to alpha-type genes. Determination of the complete sequence of 6F6.1 and of the major exon of the A-gene near it shows that these two genes have a convergent rather than a divergent direction of transcription and thus do not constitute a typical gene pair. The sequence data further suggest that 6F6.1 is transcriptionally active. The three dispersed 6F6 gene copies are localized at points of inversion of the polarity of neighboring gene pairs and their locations in this locus indicate complex gene rearrangement events.

Amino Acid Sequence↗
PubMed1992

The possible evolutionary significance of repeat elements near and within an early chorion gene in the late chorion locus of Bombyx mori.

Three beta-type early chorion gene copies (6F6.1, 6F6.2, and 6F6.3) are dispersed in the late region of chorion locus Ch1-2. Detailed analysis of the 5'-flanking region and the intron of 6F6.1 shows that they contain sequences that are homologous to Bombyx mori Bm1 repeat elements. Interestingly, the Bm1-type segment of the intron is interrupted by the insertion of a sequence that shows significant similarities with part of an intron of B. mori and Bombyx mandarina fibroin genes, and with part of the 3'-flanking region of B. mori prothoracicotropic hormone and tRNA-Glu genes; this sequence may represent a new repetitive, possibly transposable, element of B. mori. Following the Bm1-homologous sequence of the 6F6.1 5'-flanking region and preceding the gene promoter region, a short DNA segment shows sequence motifs that are also present in the ErA.1 promoter region. The occurrence of these sequences near one end or within the Bm1 repeat element is suggestive of complex sequence transfer events. Comparative analysis of known B. mori chorion alpha-gene promoters and of Bm1 repeat elements suggests, with marginal statistical significance, that these two sets of sequences contain common elements.

Animals↗
PubMed1988

Organization and expression of three genes from the silkmoth early chorion locus.

Chorion genes in the silkmoth Bombyx mori are clustered in a small region of one chromosome. Genes are grouped within this locus according to their expression in either early, middle, or late stages of choriogenesis. The entire set of late genes and a large fraction of the middle genes have been cloned and extensively characterized. We report here the first molecular characterization of a set of early chorion genes clustered within a 22.5-kb region. Transcripts homologous to these genes appear in the very first choriogenic follicle but disappear with different kinetics. One of the three early genes, 5H4, is the first example of a chorion gene which is not a member of a multigene family, or divergently paired with another chorion gene. The other two early genes, (ErA.1 and ErB.1) are divergently paired with each other in the manner of the middle and late chorion genes. The common 5' region between ErA.1 and ErB.1 is significantly shorter and does not contain certain sequence elements shared by the highly conserved 5' regions of middle and late chorion gene pairs. This evidence taken together with the fact that 5H4 is unpaired indicates that the early genes may utilize a regulatory mechanism somewhat different from the middle and late gene families.

Amino Acid Sequence↗
PubMed1986

Evolution of the silk moth chorion gene superfamily: gene families CA and CB.

The nucleotide sequences of two developmentally early chorion cDNA clones from Bombyx mori define two distinct proline-rich chorion protein families, which we name CA and CB to indicate their homologies to the previously defined chorion protein families A and B, as well as the developmentally late and cysteine-rich HcA and HcB chorion families. Thus, the chorion gene superfamily has two symmetrical branches, each consisting of three families: the alpha branch (A, CA, HcA families) and the beta branch (B, CB, HcB families). The evolution of the superfamily is discussed.

Animals↗
PubMed1985

A complex set of early chorion DNA sequences from Bombyx mori.

A follicular cDNA library from Bombyx mori (576 clones) was screened to obtain a representative sample of chorion sequences differentially expressed during the early period of choriogenesis. Ten selected sequences were characterized by Northern analysis and by dot blots of stage-specific RNA. Five of these sequences (represented in the library by 24 clones) had the developmental specificity and transcript size expected of early chorion components; two sequences (37 clones) had the developmental specificity expected of middle and late chorion components; and three sequences (16 clones) were presumed to be non-chorion, since they are constitutive or disappear at the beginning of choriogenesis. The five recovered early chorion cDNA components have been sequenced, and define three distinct subfamilies of the chorion B multigene family, and two C type families. The complexity of the early chorion genes in B. mori is discussed.

Animals↗
PubMed1984

Ribosomal DNA genes of Bombyx mori: a minor fraction of the repeating units contain insertions.

We have analyzed multiple recombinant DNA clones containing ribosomal RNA repeat units of the silkmoth, Bombyx mori. In combination with genomic DNA blots, analysis of these clones indicated that the rDNA repeat of B. mori is 10.8 kilobase pair in length and tandemly repeated in the genome, as reported by Manning et al. (18). However, contrary to that report, approximately 12% of the rDNA cistrons are interrupted by insertions of non-ribosomal DNA. Two classes of DNA insertions were identified. In one class the insertions are positioned in a region of the 28S coding sequence similar to that of the predominant rDNA insertions found in a variety of Dipteran and Tetrahymena species. In the second class, probable insertions are found close to the 3' terminus of the 28S coding sequence. Restriction enzyme analysis indicates that the two classes of insertions are not related.

Animals↗
PubMed1984

Diversity in a chorion multigene family created by tandem duplications and a putative gene-conversion event.

Two families of high-cysteine chorion proteins in Bombyx mori are encoded in 15 tandemly arranged nonidentical gene pairs. It is assumed that this locus arose by duplication with subsequent sequence divergence. We have compared DNA sequences from two such neighboring pairs of genes in an attempt to understand the manner in which diversity has been generated and/or removed. A high level of sequence identity (91%-99%) was found between the repeats throughout the transcribed and flanking regions, with two significant exceptions. First, in the DNA segment encoding a conserved region of the chorion proteins, ten substitutions were detected in a 39-base-pair region. This localized region of high variability would suggest an intergene conversion-like event. Second, a length difference of 141 base pairs was detected in a region encoding the carboxy-terminal arm of the protein. This difference can be explained by three separate reiterations of single codons (3 base pairs) separated in time by duplication or triplication events.

Amino Acid Sequence↗
PubMed1983

Novel B family sequence from an early chorion cDNA library of Bombyx mori.

A cDNA library has been constructed from transcripts present during the early stages of choriogenesis in the silk-moth, Bombyx mori. Clones representing novel sequences were selected by screening the library with (i) homotypic cDNA, (ii) rDNA and typical chorion probes of the A and B multigene families and (iii) selected clones from the library, which were positive in step (i) but negative in step (ii); this third step identified clones that hybridized to distinct subsets of the library. The sequence of one such clone was determined and shown to represent a novel subfamily of the B chorion multigene family, exclusively expressed during the early choriogenic period.

Amino Acid Sequence↗
PubMed1975

The thermodynamics and kinetics of conformational changes in 5-S RNA from Escherichia coli.

Solutions of native, renatured and denatured 5-S RNA from Escherichia coli were each found to revert to a mixture of the denatured and renatured forms when incubated at elevated temperatures in the absence of divalent cations. The equilibrium and rate constants for the renaturation and denaturation reactions were studied as a function of temperature and concentration of added NaCl. The enthalpy and entropy of denaturation and the activation energies were determined. The activation energies for the renaturation reaction were found to be almost independent of the salt concentration, whereas the other parameters increased with the salt concentration. The results are interpreted in terms of a partially unfolded intermediate whose concentration is suppressed by the addition of salt.

Binding Sites↗