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K Huppi

Publications and source records attributed to K Huppi.

At least 55 records · Page 3Linked to original sources
PubMed1992

Molecular cloning of the human brain and gastric cholecystokinin receptor: structure, functional expression and chromosomal localization.

The receptors for the brain and gastrointestinal peptide, cholecystokinin, can be classified into CCKA and CCKB subtypes. Having recently cloned the rat CCKB receptor, we used it's cDNA to isolate the human CCKB receptor homologue from brain and stomach which encodes a 447 amino acid protein with 90% identity to both rat CCKB and canine gastrin receptors. Northern hybridization identifies transcripts from stomach, pancreas, brain and gallbladder. The CCKB receptor gene maps to chromosome 11. Expression of the receptor cDNA in COS-7 cells was characteristic of a CCKB receptor subtype pharmacology. These data confirm that we have cloned a novel gene for the human brain and stomach CCKB receptor.

Amino Acid Sequence↗
PubMed1992

The pattern of p53 mutations in Burkitt's lymphoma differs from that of solid tumors.

Available evidence suggests that, among hematological malignancies, p53 is most often mutated in Burkitt's lymphoma (BL). However, much of the published data is based on cell lines. We have, therefore, analyzed BL biopsies to determine more accurately the frequency and pattern of p53 mutations in primary tumors and to determine whether there are differences among the various subtypes of BL. Among 27 BL biopsies from South Africa, we have observed mutations in the p53 gene (exons 5 through 8) in 37% of tumors. The higher frequency of mutations in cell lines (70%) suggests that mutation of p53 may be associated with tumor progression. Summarizing available data we conclude that the presence of mutated p53 in BL is independent of the geographic origin of the tumor, the 8;14 chromosomal breakpoint locations and Epstein-Barr virus association. We also find that the mutational spectrum of p53 in BL differs from that observed in nonlymphoid tumors. More than 50% of mutations in BL are clustered in a small stretch of 33 amino acids (codons 213 to 248). Interestingly, codon 213 appears to be as frequently mutated as codon 248. Conversely, codon 273, often mutated in solid tumors, is rarely involved in BL.

Amino Acid Sequence↗
PubMed1992

Infrequent p53 mutation in mouse tumors with deregulated myc.

An invariant genetic lesion in mouse plasmacytomas is deregulated expression of c-myc as a consequence of chromosomal translocation. However, retroviral and transgenic studies suggest that additional genetic lesions may contribute to the genesis of plasmacytomas. The p53 tumor suppressor gene is a likely contributor to this genetic lesion, since there is a high incidence of p53 mutation in Burkitt's lymphomas and B-ALL (L3), both of which contain translocations involving c-myc analogous to those in plasmacytomas. In addition, p53 has been shown to be a transcriptional modulator of c-myc expression. In a survey of 27 mouse plasmacytomas by single-strand conformation polymorphism, we identified a single mutation (3.7% incidence), suggesting that p53 lesions are not frequent contributors to plasmacytomagenesis. A similar study of macrophage-monocyte tumors generated by a c-myc-containing retrovirus also indicates a lack of p53 involvement in deregulated c-myc expression. These results suggest that the specific maturation stage of transformed B-lymphocytes, independent of c-myc deregulation, may be the critical factor which determines the involvement of mutant p53.

Amino Acid Sequence↗
PubMed1992

Expression of murine cyclin B1 mRNAs and genetic mapping of related genomic sequences.

Two cDNAs that encode a protein with 87% identity to human cyclin B1 and that differ only in the length of their 3'-untranslated regions have been isolated from a 70Z/3B murine pre-B leukemia cell library. Three sizes of RNA transcripts were detected in Northern hybridization analyses of a variety of normal tissues and transformed cell lines using the cDNA inserts as probes. The expression of these RNAs can be modulated in tissue culture cell lines by physiologically relevant stimuli, increasing when cells are stimulated to proliferate and decreasing when cells are induced to differentiate. Moreover, RNAs from tissues that contain few proliferating cells have no detectable hybridizing transcripts. The coordinate regulation of these RNAs with other genes that are activated during the cell division cycle and the profound similarity of the predicted amino acid sequence to those of published cyclin B homologues indicate that these genes encode a murine cyclin B1. In Southern hybridization analysis of BALB/cAnPt genomic DNA digested with EcoRI, 12 fragments hybridized with the cDNA probes. Through Southern blot analyses of DNA from backcross and cogenic mice, recombinant inbred strains, and somatic cell hybrids, the genetic loci that produce the cyclin B1-related sequences (designated loci Cycb1-rs1 to Cycb1-rs9) were mapped on mouse chromosomes 5, 1, 17, 4, 14, 13, 7, X, and 8, respectively. Cycb1-rs6 (on chromosome 13) is discussed as the most likely candidate for an expressed structural gene locus.

3T3 Cells↗
PubMed1991

Mouse bcl-3: cDNA structure, mapping and stage-dependent expression in B lymphocytes.

Human B-cell chronic lymphocytic leukemias (CLLs) are malignancies of mature B lymphocytes. A subset of these tumors is associated with a non-random t(14; 19) translocation (Ueshima et al., 1985). Recently a gene (bcl-3) has been identified in the region adjacent to the chromosome 19 breakpoint in this translocation (McKeithan et al., 1987; Ohno et al., 1990). We now report the isolation of cDNA clones of mouse bcl-3. The mouse bcl-3-coding region is 1746 bp long and exhibits 80% identity with human bcl-3 at both the nucleotide and amino acid level. The bcl-3 locus maps to the proximal end of mouse chromosome 7, which is syntenic to human chromosome 19. The bcl-3 probe readily detects particularly abundant amounts of a 1.8 kb mRNA in mouse tumors consisting of follicular center mature B cells and large pre-B cells, but not in small pre-B cells. The bcl-3 pattern of expression is distinctive in the spectrum of B-cell maturation in that bcl-3 transcripts are particularly abundant in B-cell lines immortalized just prior to Ig switch. The bcl-3 pattern of expression also bears close resemblance to that of bcl-2 (Gurfinkel et al., 1987), which is frequently associated with human B follicular lymphomas [t(14; 18)] and some chronic lymphocytic leukemias (Adachi et al., 1989; 1990; Adachi & Tsujimoto, 1989).

Amino Acid Sequence↗
PubMed1990

A deletion linked to a poly(ADP-ribose) polymerase gene on chromosome 13q33-qter occurs frequently in the normal black population as well as in multiple tumor DNA.

The nuclear enzyme poly(ADP-ribose) polymerase (PADPRP) is thought to play a role in DNA recombination, replication, and repair. In view of the implication of these processes in tumorigenesis, and based on preliminary evidence which indicated the presence of an extraneous polymorphic restriction fragment for murine PADPRP loci in strains of mice susceptible to plasmacytomas, we investigated correlations between the restriction fragment length polymorphism of the PADPRP gene(s) and human Burkitt lymphoma. No increase in the frequency of polymorphisms on chromosome 1 (containing the active gene) or on chromosome 14 (a pseudogene) was observed. However, restriction fragment length polymorphism analysis of PADPRP sequences on chromosome 13 (either a processed pseudogene or a gene with extensive identity to PADPRP) revealed that of 19 DNA samples derived from endemic Burkitt lymphoma all contained at least one copy of a rare allele (B). Simple two-allele (A/B) polymorphisms in this PADPRP-like locus were identified by digestion with a number of restriction enzymes including HindIII, PstI, KpnI, and MspI. These restriction fragment length polymorphisms always segregated together, suggesting that they identify a deletion within or close to the PADPRP sequences on chromosome 13, which we mapped precisely to 13q33-qter. Based upon family studies the A and B alleles were shown to be transferred in a Mendelian codominant fashion. Subsequently, this probe was used as a linkage marker to study the frequency of this deletion in various tumors including B-cell follicular lymphomas, small cell lung carcinomas, breast carcinomas, and colorectal carcinomas. In noncancer control populations, the frequency of this deletion was 3-fold higher among Blacks as compared to Caucasians. When DNA from various tumors was compared to normal DNA from racially appropriate noncancer controls, the frequency of this deletion was still 2- to 3-fold higher in the tumor DNA. Matched samples provided instances of tumor-specific loss of heterozygosity but also revealed that the predominant source of this deletion is the germ line, suggesting that the chromosome 13 region neighboring the PADPRP locus may harbor a gene whose loss may predispose individuals to malignancy.

Alleles↗
PubMed1990

Cell cycle regulation of an exogenous human poly(ADP-ribose) polymerase cDNA introduced into murine cells.

We have evaluated the regulation of expression of the poly(ADP-ribose) polymerase gene during cell growth and replication. In a synchronized population of HeLa cells or in serum-stimulated WI-38 cells, steady-state levels of the polymerase mRNA were highest at late S and S-G2 phases and negligible in early S phase. Transcription did not solely account for the significant increase in the mRNA levels observed in late S phase by Northern analysis. The stability of the mRNA was dependent upon the percent proliferating cells in the culture. Accordingly, polymerase mRNA from cells in early exponential phase was significantly more stable than from cells in stationary phase of asynchronous growth. To clarify these observations, we utilized a novel heterologous expression system that involved murine 3T3 cells transfected with a human poly(ADP-ribose) polymerase cDNA under the control of a non-cell cycle-specific promoter. Cells were synchronized, and a comparison was made of the endogenous (murine) and exogenous (human) polymerase mRNA levels. Both the endogenous and the exogenous mRNA were specifically stabilized by the same mechanisms and only during late S phase; therefore, we concluded that mRNA pools for the polymerase are regulated at the post-transcriptional level. The heterologous expression system confirmed that the post-transcriptional regulation system in the mouse cells can recognize and faithfully regulate the human cDNA in response to the murine cell cycle signals. More importantly, the presence of extra copies (human) of the polymerase gene did not provide an increased amount of the total polymerase mRNA or protein and, in fact, the sum of the endogenous and exogenous mRNA in the transfected cells was approximately the same as the level of endogenous transcript in the control cells. This suggested that there might be a limit to the amount of polymerase protein accumulating in the cellular pool and thus levels of poly(ADP-ribose) polymerase may be autoregulated.

Animals↗
PubMed1990

Pvt-1 transcripts are found in normal tissues and are altered by reciprocal(6;15) translocations in mouse plasmacytomas.

The mouse Pvt-1 (for plasmacytoma variant translocation) region maps to a chromosome 15 breakpoint region that is frequently interrupted by "variant" reciprocal chromosome translocations, rcpt(6;15), in plasmacytomas. This region lies several hundred kilobases (kb) 3' of the mouse c-myc gene (Myc) which is deregulated in both rcpt(6;15) and rcpt(12;15) plasmacytomas. rcpt(12;15) translocations apparently activate c-myc directly by interrupting the gene itself, but the mechanism causing c-myc deregulation in tumors bearing rcpt(6;15) translocations remains unknown. The indirect activation of c-myc by Pvt-1 interruption has remained an appealing possibility, but heretofore it has not been possible to establish such a connection. Furthermore, no genes from the Pvt-1 locus have been shown to be transcribed in normal tissues or in tumors with rcpt(6;15) translocations. We report the isolation of a cDNA clone, Pvt-1-1, from mouse spleen mRNA that is specific to the Pvt-1 region. This cDNA probe detects low levels of large (ca. 14 kb) RNA transcripts in normal mouse tissues. In plasmacytomas with rcpt(6;15) translocations, the Pvt-1 transcripts are elevated in abundance and truncated in size. Both changes are apparently induced by the chromosomal translocation. Expression of 14-kb Pvt-1 RNA is elevated in B-cell tumor lines that express immunoglobulin light chain genes; thus, we postulate that these translocations are facilitated by the increased DNA accessibility of immunoglobulin kappa light chain and Pvt-1 genes when they are simultaneously expressed at certain times during B-cell ontogeny.

Amino Acid Sequence↗
PubMed1989

Sequence and organization of the mouse poly (ADP-ribose) polymerase gene.

Using a human cDNA probe, we have isolated murine genomic and cDNA clones corresponding to the nuclear enzyme poly (ADP-ribose) polymerase (ADPRP). Northern analysis with the mouse cDNA clones reveals transcripts of 3.7-3.8 kb corresponding in size to the human ADPRP transcript. DNA sequence comparisons between mouse and human clones reveals extensive amino acid sequence conservation within regions harboring DNA binding, NAD+ binding or automodification domains. A survey among mouse inbred strains for restriction fragment length polymorphism (RFLP) reveals at least three distinct ADPRP alleles. The segregation of alleles among mouse genetic recombinants positions ADPRP on mouse chromosome 1 between the complement receptor-related gene At-3 and the Fc receptor locus FcR. Furthermore, ADPRP is closely associated with the autoimmune locus gld (generalized lymphadenopathy).

Amino Acid Sequence↗
PubMed1988

Receptors for Fc epsilon and Fc gamma are linked on mouse chromosome 1.

Recently isolated cDNA clones for the high affinity Fc epsilon receptors on mast cells and basophils (Fc epsilon RI alpha) and Fc gamma receptors on macrophages and lymphocytes (Fc gamma 2b/gamma 1R) are homologous members of the Ig supergene family. Analysis of the segregation of restriction fragment length polymorphism in crosses of inbred mice now establish that the structural genes encoding both Fc epsilon RI alpha and Fc gamma 2b/gamma 1R are indeed discrete genes and are linked at the distal end of mouse chromosome 1. This finding raises the possibility that a family of Fc receptors could be found in a region that is known to contain immunologically important markers of lymphocyte surface Ag and autoimmune defects.

Animals↗