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Biomedical subjects

P Gerlinger

Publications and source records attributed to P Gerlinger.

At least 37 records · Page 2Linked to original sources
PubMed1988

Ha-ras and c-myc oncogene expression interferes with morphological and functional differentiation of mammary epithelial cells in single and double transgenic mice.

We studied the effects of the tissue-specific and lactogenic hormone-dependent expression of the recombinant whey acidic protein (Wap)-ras and Wap-myc oncogenes on the differentiation of the mammary epithelium in transgenic mice. The histological appearance of mammary glands in pregnant, lactating, and postlactational animals and their ability to express milk protein genes were analyzed. Activated Ha-ras expression caused a decrease of milk protein synthesis during the lactation period. The formation of glandular epithelium and the postlactational regression of epithelial cells were not affected. c-myc expression impaired the development of the glandular epithelium, and milk protein synthesis was decreased strongly. Epithelial cell proliferation continued during lactation and postlactationally. Coexpression of both oncogenes in double transgenic mice synergistically affected differentiation and resulted in a high number of neoplastic foci. Palpable tumors were observed only after a latency of 3-4 months. Tumor cells utilize the Wap promoter hormone independently, express increased levels of Wap-ras and induce adjacent stromal cells to produce tenascin.

Animals↗
PubMed1988

B-cell control region at the 5' end of a major histocompatibility complex class II gene: sequences and factors.

Transcription of major histocompatibility complex class II genes is elaborately regulated. Mouse class II genes are transcribed primarily in B cells, peripheral macrophages and interdigitating cells, and thymic cortical and medullary cells. In this study, we began to identify the DNA sequences and protein factors that control expression of a class II gene in B cells, addressing in particular how closely they resemble those that regulate immunoglobulin gene expression. We describe a region upstream of the E alpha gene that is crucial for its transcription in the B cells of transgenic mice but is less important in cultured B-cell lines. The sequence of this region reveals several familiar motifs, including a second X-Y pair reminiscent of that residing in the promoter-proximal region of all class II genes, a B motif strikingly homologous to that associated with the immunoglobulin kappa gene enhancer, several Ephrussi motifs, and a Pu box-like sequence very similar to that implicated in simian virus 40 and lymphotrophic papovavirus expression in B cells. Careful study of the proteins that bind specifically to these different motifs prompts us to suggest that major histocompatibility complex class II and immunoglobulin genes rely on quite different factors to achieve B-cell-specific expression.

Animals↗
PubMed1987

Ha-ras oncogene expression directed by a milk protein gene promoter: tissue specificity, hormonal regulation, and tumor induction in transgenic mice.

The activated human Ha-ras oncogene was subjected to the control of the promoter region of the murine whey acidic protein (Wap) gene, which is expressed in mammary epithelial cells in response to lactogenic hormones. The Wap-ras gene was stably introduced into the mouse germ line of five transgenic mice (one male and four females). Wap-ras expression was observed in the mammary glands of lactating females in two lines derived from female founders. The tissue-directed and hormone-dependent Wap expression was conferred on the Ha-ras oncogene. The signals governing Wap expression are located within 2.5 kilobases of 5' flanking sequence. The other two lines derived from female founders did not express the chimeric gene. In the line derived from the male founder, the Wap-ras gene is integrated into the Y chromosome. Expression was found in the salivary gland of male animals only. After a long latency, Wap-ras-expressing mice developed tumors. The tumors arose in tissues expressing Wap-ras--i.e., mammary or salivary glands. Compared to the corresponding nonmalignant tissues, Wap-ras expression was enhanced in the tumors.

Animals↗
PubMed1986

B-lymphocyte targeting of gene expression in transgenic mice with the immunoglobulin heavy-chain enhancer.

A hybrid gene containing rabbit beta-globin structural sequences (-9 to +1650), and a chicken conalbumin gene promoter (+62 to -102) in the place of the beta-globin promoter (upstream from -9), was inactive in 5 different transgenic mouse line. Adding the mouse immunoglobulin heavy-chain (IgH) enhancer to this construction specifically stimulated expression in B-cells. These results show that IgH enhancer is specifically active in B-cells. Expression of the hybrid gene was low compared to the endogenous immunoglobulin heavy and light-chain genes. Substituting the mouse immunoglobulin kappa light-chain gene (Ig kappa) promoter (+4 to -800) for the heterologous conalbumin promoter was not sufficient to restore gene expression to level of the endogenous genes. In addition to the reproducible B cell expression, we also found inheritable unexpected expression in certain tissues, which varied from line to line.

Animals↗
PubMed1985

Transvaginal sonographically controlled follicle puncture for oocyte retrieval.

Transvaginal sonographically guided follicle aspiration under local anesthesia was performed on more than 100 patients and compared favorably with the other techniques that have been proposed to retrieve oocytes for in vitro fertilization. Sonography employs a sector scanner placed on the abdomen, and the needle is introduced through the posterior fornix of the vagina into the cul-de-sac and the ovary. In case of a high ovary, a transvaginal-transvesical variation may be used. The method is not painful and is easy to learn and perform. The sole adverse incidents have involved inadvertent venous puncture with no sequelae. The number and quality of recovered oocytes are good, and five normal children conceived after this method of retrieval were recently born. The technique permits substantial simplification of egg recovery for in vitro fertilization, which can now be performed in an outpatient setting without the risk and expense of laparoscopy and general anesthesia or the discomfort of transabdominal-transvesical ultrasound-guided aspiration.

Clomiphene↗
PubMed1984

Termination of the ovalbumin gene transcription.

RNA transcribed in vivo from the chicken ovalbumin gene has been analyzed in detail in the 3' region of the gene using nuclease S1 mapping and Northern blotting. We describe two new additional minor polyadenylation sites leading to pre-messenger RNA with lengths of approximately 7900 and 9700 nucleotides. Hybridization of RNA transcribed in vitro from oviduct nuclei with various immobilized DNA fragments covering the 3' region of the gene indicates that greater than 90% of transcription terminates in a discrete region of 170 bp located approximately 900 bp downstream from the 3' end of the last exon. Two copies of a sequence homologous to the one proposed for yeast transcription termination are present within the region where transcription of the ovalbumin gene terminates.

Animals↗
PubMed1981

The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation.

The ovalbumin gene family is composed of three genes, X, Y and ovalbumin, which are expressed in laying hen oviduct. We have analyzed the in vivo transcription products of X and Y genes and the effect of steroid hormones on their synthesis and accumulation. As in the case of ovalbumin, the complete gene transcripts and processing intermediates are present in the poly(A)+ RNA fraction. The mature RNAs are found in polysomes and are translated into proteins. The expression of X and Y genes is controlled by steroid hormones: X and Y RNAs are not detectable in oviducts from chicks withdrawn from estrogen stimulation, whereas in chicks stimulated with estrogen for 7 days, X RNA represents 0.3% and Y RNA 0.8% of ovalbumin mRNA. In laying hen, however, the levels of X and Y RNAs are about 2% of ovalbumin mRNA. After stimulation with other steroid hormones, alone or in combination, the level of X and Y RNA does not achieve that detected in laying hen. Progesterone has a much weaker effect on X RNA accumulation than on that of Y and ovalbumin mRNAs. Studies with isolated nuclei show that X and Y gene expression is regulated by hormones at the level of transcription. However, the differences observed between the transcription rates and the accumulation of X and Y mRNAs suggest that the expression of X and Y genes could also be controlled at the levels of RNA processing and/or mRNA stability.

Animals↗
PubMed1979

The chicken conalbumin gene: studies of the organization of cloned DNAs.

The construction of a double-stranded conalbumin cDNA plasmid (1) has allowed us to investigate the structure of the conalbumin gene. Restriction enzyme mapping of chicken genomic DNA reveals that the conalbumin gene is split and is contained in three EcoRI fragments "a", "b", "b" and "c" which have sizes of 10.7, 4 and 2.5 kb, respectively. Analysis with specific probes shows that the orientation of these fragments with respect to transcription is 5'-"b", "c" and "a"-3. The fragments Eco "b", Eco "c" and part of Eco "a" have been isolated by molecular cloning from three different "libraries". Electron microscopic studies of hybrids between cloned DNA's and conalbumin mRNA show that one of the isolated clones, lambda C4-conl, contains the coding sequences for the first 940 nucleotides of the mRNA (out of 2400). This region is highly split, since it contains seven short exonic sequences separated by six intervening sequences. The DNA region coding for these 940 nucleotides is 5 times longer than the mRNA coding sequences, a ratio very similar to that found for other chicken genes.

Animals↗
PubMed1978

Nucleotide sequence at the 5' extremity of tobacco-mosaic-virus RNA. 2. The coding region (nucleotides 69-236).

In the preceding paper it was shown that the first A-U-G codon in tobacco mosaic virus RNA is separated from the 5' terminus by a sequence of 68 nucleotides devoid of internal guanosine residues. In this paper we present the sequence of 165 residues immediately following the first potential initiation codon. The characterized sequence contains four nonsense codons but none are in phase with the prospective initiation codon. Several lines of evidence, including direct characterization of the portion of the RNA molecule which binds to and is protected by the ribosome in the course of initiation, all support the idea that the A-U-G at position 69-71 is a functional initiation signal for viral protein synthesis.

Base Sequence↗
PubMed1978

Electron microscopy and restriction enzyme mapping reveal additional intervening sequences in the chicken ovalbumin split gene.

The Eco RI fragment "b" of chicken DNA (Breathnach, Mandel and Chambon, 1977), which contains the sequences coding for the 5' quarter of ovalbumin mRNA (ov mRNA), has been isolated by molecular cloning using a "shotgun" approach. Electron microscopy and restriction enzyme analysis have revealed that the sequences coding for the 5' quarter (approximately 500 nucleotides) of ov mRNA are split into four regions separated by three intervening sequences. The cloning procedure seems to be reliable, since the restriction enzyme pattern of the cloned Econ RI fragment "b" is similar to that of the corresponding chromosomal DNA fragment. There is no evidence supporting the existence of a 150-200 nucleotide long sequence at the 5' end of the ov mRNA similar to the "leader" sequences found at the 5' end of some adenovirus and SV40 mRNAs.

Animals↗
PubMed1978

Organization of coding and intervening sequences in the chicken ovalbumin split gene.

The interruptions in the chicken ovalbumin gene which were reported previously (Breathnach, Mandel and Chambon, 1977) are shown to be due to the presence of intervening sequences which separate the messenger-coding sequences. We present evidence for an additional interruption of the gene, which, together with those reported earlier and by Garapin et al. (1978b), make a total of six intervening sequences. All of these intervening sequences are located in the DNA region that corresponds to the part of the ov mRNA which codes for amino acids. The seven coding fragments of the split ovalbumin gene are arranged in the same order and relative orientation as in the ovalbumin double-stranded cDNA. All the sequences coding for ov mRNA are contained in a chromosomal DNA region of 6000 bp, which is more than 3 times longer than ov mRNA. The general organization of the ovalbumin split gene is discussed.

Animals↗
PubMed1978

The ovalbumin split gene: molecular cloning of Eco RI fragments "c" and "d".

The Eco RI fragments "c" and "d" of the ovalbumin gene (1, 2) have been isolated by molecular cloning. Restriction enzyme mapping and electron microscopy have confirmed that the two fragments contain the same ovalbumin mRNA coding sequences. These sequences are split into two regions which have been mapped in fragments "c" and "d". There is no evidence that the ovalbumin mRNA sequences contained in these fragments could be further interrupted. Our results confirm that the presence of Eco RI fragment "d" in some chickens is due to the existence of an allelic variant of the ovalbumin gene which contains an additional Eco RI site within the region corresponding to Eco RI fragment "c". This additional Eco RI site appears to be the main difference between the two alleles. Finally, our results provide a direct demonstration that most of the ovalbumin mRNA sequences are encoded for by Eco RI fragments "a", "b" and "c".

Coliphages↗
PubMed1977

Monocistronic translation of alfalfa mosaic virus RNAs.

The four alfalfa mosaic virus RNAs (respectively 24 S, 20 S, 17 S and 12 S) have been used separately as messengers in two in vitro protein synthesizing systems: wheat germ and rabbit reticulocyte lysate. In both systems a polypeptide corresponding to the translation of the entire length of the RNA can be found for RNAs 24 S, 20 S and 12 S, but not for 17 S RNA, the translation product of which is only 35,000 daltons. The number of initiation sites has been determined for each RNA by analyzing the initiation peptides synthesized in the presence of spasomycin and show that there is only one initiation or binding site perRNA. We thus conclude that each AMV RNA behaves as a monocistronic messenger in in vitro translating systems.

Animals↗
PubMed1977

Molecular cloning of extensive sequences of the in vitro synthesized chicken ovalbumin structural gene.

Double-stranded DNA molecules complementary to ovalbumin chicken messenger RNA were synthesized in vitro and integrated into the E. coli plasmid pCR1 using an oligodG-dc tailing procedure. The resultant hybrid plasmids, amplified by transfection of E. coli, were shown by hybridization and gel electrophoresis to contain extensive DNA sequences of the ovalbumin structural gene.

Animals↗