Biomedical subjects

S Mazan

Publications and source records attributed to S Mazan.

30 records · Page 2Linked to original sources

PubMed1993

In humans all U3 genes map to chromosome 17p12-->p11, but in mouse the U3A and U3B genes are located on different chromosomes.

U3 small nucleolar RNA, which participates in eukaryotic rRNA processing, is encoded by a small multigene family in mammals. In humans, the four to six gene copies code for an identical U3 RNA molecule; rodents, however, have two variant forms of these genes, U3A and U3B. We show that all U3 genes in humans map to a single chromosomal locus, 17p12-->-p11, which corresponds exactly to the region of mouse Chromosome 11 where the four U3B genes are clustered. By contrast, in mouse the unique U3A gene copy is not linked to the U3B gene cluster but maps to another chromosome (the B2-B4 region of Chromosome 10).

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PubMed1992

Structure of the differentially expressed mouse U3A gene.

Two markedly different forms of U3 RNA are present in mouse, the relative abundance of which largely depends upon the tissues. In all cases studied so far, the more abundant form is U3B, encoded by four previously characterized genes. We report here the isolation and analysis of the unique gene encoding the U3A variant, which completes the characterization of the mouse U3 multigene family. Comparisons with rat U3 genes indicate that the diversification of the A and B forms has predated the mouse/rat separation. The two forms of U3 RNA are submitted to similar, but not identical, primary and secondary structure constraints. As for the sequences flanking the RNA coding region, similar observations emerge for both types of genes: for each type, the 5' flanks are strongly conserved between mouse and rat, over at least the proximal 500 bp, whereas only about 30 bp of proximal 3' flanks are preserved, which include a signal for the formation of vertebrate U small nRNA 3' end. By contrast the 5' flanks of the two types of genes diverge extensively from each other, either in mouse or in rat, and could be involved in the differential expression of the two forms. Even over the few conserved motifs thought to be involved in the basic transcriptional control of vertebrate U small nRNA genes, the A and B forms of U3 genes exhibit specific differences maintained in the two rodent species.

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PubMed1992

Alterations of nucleolar ultrastructure and ribosome biogenesis by actinomycin D. Implications for U3 snRNP function.

We have studied, at the electron microscope level, the reorganizations of nucleolar ultrastructure induced by actinomycin D (AMD) in different conditions of drug treatment associated with an inhibition of rRNA synthesis. We have analyzed in parallel the localizations of ribosomal genes, of their transcripts, of various pre-rRNA intermediates, as well as of U3 RNA and fibrillarin by in situ hybridization with nucleic acid probes and immunocytological detection on thin sections of human and mouse cells. Consistent with previous observations, dense fibrillar component (DFC) and granular component (GC) appear to contain distinct pre-rRNA species at different stages of their processing. DFC appears as a major site of U3 RNA accumulation, but a very substantial fraction of nucleolar U3 RNA is also found in GC, colocalizing with partially processed pre-rRNAs. Remarkably, the major nucleolar components retain their ultrastructural appearance when extensively depleted of their pre-rRNA moiety, and ribosomal genes are always detected over fibrillar center (FC), even after extended AMD treatments which result in the characteristic segregation of nucleolar components. Moreover, while for GC the U3 RNA and pre-rRNA contents evolve in parallel following the cessation of rRNA synthesis, a dramatic uncoupling is observed for DFC. The persistent presence of U3 RNA and fibrillarin after pre-rRNA depletion suggests that DFC could represent an anchorage site for U3 snRNPs, before their entering another cycle of pre-rRNA processing reactions.

3T3 Cells↗

PubMed1991

Composition and chromosomal localization of the small multigene family encoding mouse U3B nucleolar RNA.

U3 small nucleolar RNA, which is believed to play a role in eukaryotic rRNA processing, is encoded by a small family of genes (5-10 copies/haploid genome) in mammals. In mouse, functional genes encoding the major U3B RNA form have been isolated, with all copies identified so far having evolved in a tightly concerted fashion. However, knowledge of the precise number and relative localization of all gene-family members has been hampered by the presence of multiple copies of U3B-processed pseudogenes in the mouse genome. In this study, we took advantage of a probe that is specific for functional U3B genes to address this question, using both Southern hybridization of genomic DNA and in situ hybridization of metaphase chromosomes. We show that the mouse haploid genome contains four functional U3B genes, all of which are clustered in a single chromosomal locus. They map to the C-D bands of mouse chromosome 11, within one of the most extended segments of gene-linkage conservation known between the mouse and human genomes, corresponding to a major portion of human chromosome 17. By contrast, the multiple (nonfunctional) U3 retrogenes are dispersed over several mouse chromosomes.

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PubMed1991

Localization of U3 RNA molecules in nucleoli of HeLa and mouse 3T3 cells by high resolution in situ hybridization.

We have examined the ultrastructural localization of U3 RNA in the nucleoli of HeLa and mouse 3T3 cells by in situ hybridization with a biotinylated U3 DNA probe and subsequent detection of hybrids with electron microscopy by direct immunogold labeling. The highest levels of signal density for U3 RNA are detected over the dense fibrillar component (DFC) of the nucleolus, including the interfaces between DFC and the enclosed fibrillar center (FC) on the one hand and DFC and the granular component (GC) on the other hand. Lower but significant signals also are observed over GC, which indicate, taking into account the high relative volume of GC in a nucleolus, that a substantial fraction of U3 RNA is present in this compartment where the more mature forms of pre-rRNA accumulate. In parallel, the localization of fibrillarin was analyzed by immunogold detection, demonstrating that fibrillarin and U3 RNA have a roughly similar distribution, although quantitative measurements reveal that the signal ratio for both molecules exhibit significant differences among the major ultrastructural components of the nucleolus.

3T3 Cells↗

PubMed1990

Organization of the gene family encoding mouse U3B RNA: role of gene conversions in its concerted evolution.

In mouse, U3B small nuclear RNA is encoded by a small family of four functional genes. Three of these genes have recently been identified, with two of them being located 5 kb apart, in the same orientation [Mazan and Bachellerie, J. Biol. Chem. 263 (1988) 19461-19467]. By analyzing a cosmid library of mouse genomic DNA we have isolated the fourth gene, U3B.4, which is located 12 kb downstream from the previously reported pair of linked genes, U3B.1 and U3B.2, in the same orientation. Its sequence analysis demonstrates that four mouse U3B genes have undergone a very efficient concerted evolution, with a perfect sequence identity not only over the entire RNA coding region but also over 254 bp of 5' flanking DNA which contain essential transcription signals. Moreover, in the regions which immediately flank the domain of perfect sequence identity, a patchwork of sequence similarities among the four genes is apparent, with a series of striking reversals in relationships between adjacent localized areas. This suggests that gene conversions have played a major role in the concerted evolution of the U3B gene family. A superimposition of conversion events, which probably occurred at distinct stages of rodent evolutions can be inferred. This has involved not only the three clustered copies, but also the remaining gene, U3B.3, the location of which relative to the cluster remains unknown. B1 repetitive elements are found at the boundaries of conversion domains, suggesting their possible role in control of the sequence homogenization process.

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PubMed1990

Presence of a differentially expressed U3A RNA variant in mouse. Structure and evolutive implications.

A U3 RNA variant has been identified in mouse, the abundance of which relative to the previously characterized major form (U3B) appears to vary to a large extent depending upon the cell origin. Its partial sequence analysis shows that it is clearly related to the U3A form previously described in rat. Sequence comparisons suggest that the separation of the two forms of U3 genes now found in rat and mouse represent a relatively ancient event in rodent evolution. While mouse U3B RNA is encoded by four clustered genes, the U3A variant is encoded by a unique gene. Both mouse U3 RNAs differ substantially in primary structure (more than 10% divergence). Although rodent U3 RNAs exhibit a largely similar secondary structure, a specific difference between the A and B form can nevertheless be observed.

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PubMed1989

Structure of the 5'-external transcribed spacer of the human ribosomal RNA gene.

We report the complete nucleotide sequence of the 3627 bp long 5'-external transcribed spacer (ETS) of a human ribosomal RNA gene. This sequence exhibits only very limited homologies with its mouse counterpart, the only other mammalian specimen analyzed so far. It has very peculiar compositional characteristics, with a highly biased base content (very rich in G + C, very poor in A) and also some very strong dinucleotide preferences. Interestingly, these specific features are shared by the mouse sequence, despite the extensive sequence divergence, and also apply to the other transcribed spacers of mammals indicating that a common and strong structural constraint is exerted on all these regions of the ribosomal gene. An outstanding secondary structure can be formed within the human ETS RNA, which could have a significant role in preribosome assembly.

Animals↗

PubMed1989

Mouse U3-RNA-processed pseudogenes are nonrandomly integrated into genomic DNA. Implications for the process of retrogene formation.

We have characterized three mouse U3-RNA-processed pseudogenes. Together with previous analyses of mouse functional genes and of rat genes and pseudogenes, these data provide some insights into the processes of U3 retrogene formation during the evolution of rodents. All the mouse retrogenes correspond to a full-length U3B-coding sequence with a 3'-poly(A) tail and are precisely flanked by a pair of direct repeats, in agreement with formation through an RNA intermediate followed by insertion at staggered nicks in the genome. All the rodents U3 retrogenes identified so far are produced from a U3B-RNA form, with two of them (one for each rodent) formed from a 3'-elongated U3-RNA precursor, pointing to the particular susceptibility of RNA precursors forms to serve as templates for retrogene formation. Rodent full-length U3 retrogenes are not inserted randomly in the genome but are systematically found in a context of simple sequences, prone to slipped-strand mispairings and likely to favour the appearance of single-stranded DNA. Moreover their flanking repeats share not only the same size (15 bp) but also common sequence features (which extend to vicinal upstream nucleotides) suggesting that common mechanisms, specific to this class of retrogenes, have been involved in their formation. For the first time, a model accounting for the generation of full-length cDNA copies from nonpolyadenylated RNA templates is proposed. Rodent U3 retrogenes appear to be of rather recent origin (posterior to the mouse/rat divergence) and some of them could have undergone subsequent genetic exchanges with the functional genes.

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PubMed1988

Structure and organization of mouse U3B RNA functional genes.

We report the isolation and primary structure of three genes encoding mouse U3B RNA which are expressed after injection into Xenopus laevis oocytes. Over the U3B RNA coding region, their sequences are perfectly identical, showing nine point differences with rat U3B, which do not alter the RNA secondary structure. A comparison of the three mouse sequences for the gene flanks reveals the extensive divergence of the downstream regions, except for a few nucleotides adjacent to the U3 RNA coding region, which contain a motif matching the consensus sequence for the U small nuclear RNA 3' end formation signal. By contrast, the upstream flanking regions are strongly homologous up to position -500, but they completely diverge thereafter. Within the homologous portion of 5' flanks, several motifs can be recognized which are unambiguously related to sequence elements involved in the transcriptional control of other U small nuclear RNA genes: two of these motifs precisely map at the locations (relative to the transcription start site) expected for the proximal and distal (enhancer-like) sequence elements of U small nuclear RNA genes, and "Sp1"-GC boxes and a CCAAT box are also present in their vicinity. The comparison with the rat U3B gene confirms that the preferential preservation of the 5'-flanking sequences extends up to position -500, suggesting the functional importance of sequences well upstream from the distal sequence element of the promoter. Two of the mouse genes are closely linked in genomic DNA (5 kilobase pairs apart, same orientation) and seem to have been homogenized through a recent conversion event. More generally, this small multigene family (at most six to seven copies of functional U3B genes per mouse haploid genome) appears to have undergone a concerted evolution in rodents.

Animals↗

PubMedDate not supplied

Pax6 expression patterns in Lampetra fluviatilis and Scyliorhinus canicula embryos suggest highly conserved roles in the early regionalization of the vertebrate brain.

We report expression patterns of the Pax6 gene in the dogfish Scyliorhinus canicula and the lamprey Lampetra fluviatilis during neurulation and at the beginning of organogenesis. At the stages studied, both genes display very similar expression domains in the dorsal forebrain, with a sharp posterior boundary at the diencephalon-mesencephalon border, in the hindbrain, excluding the floor plate and the roof plate, and in the spinal cord. The comparison of these expression patterns with those reported in osteichthyans suggests that the roles played by Pax6 in early brain regionalization have been highly conserved during vertebrate evolution.

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PubMedDate not supplied

Otx1 gene-controlled morphogenesis of the horizontal semicircular canal and the origin of the gnathostome characteristics.

The horizontal semicircular canal of the inner ear is a unique feature of gnathostomes and is predated by the two vertical semicircular canals, which are already present in lampreys and some fossil, armored jawless vertebrates regarded as close relatives of gnathostomes. Inactivation in mice of the orthodenticle-related gene Otx1 results in the absence of this structure. In bony fishes and tetrapods (osteichthyans), this gene belongs to a small multigene family comprising at least two orthology classes, Otx1 and Otx2. We report that, as in the mouse, xenopus and zebrafish, Otx1- and Otx2-related genes are present in a chondrichthyan, the dogfish Scyliorhinus canicula, with an Otx1 expression domain in the otocyst very similar to those observed in osteichthyans. A strong correlation is thus observed in extant vertebrates between the distribution of the horizontal semicircular canal and the presence of an Otx1 ortholog expressed in the inner ear, which supports the hypothesis that the absence of this characteristic in Otx1-/- mice may correspond to an atavism. The same conclusion applies to two other gnathostome-specific characteristics also deleted in Otx1-/- mice, the utriculosaccular duct and the ciliary process. Together with functional analyses of Otx1 and Otx2 genes in mice and comparative analyses of the Otx gene families characterized in chordates, these discoveries lead to the hypothesis that some of the anatomic characteristics of gnathostomes have appeared quite suddenly and almost simultaneously in vertebrate evolution, possibly as a consequence of gene functional diversifications following duplications of an ancestral chordate gene.

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