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Biomedical subjects

E Canaani

Publications and source records attributed to E Canaani.

At least 19 recordsLinked to original sources
PubMed1992

The t(4;11) chromosome translocation of human acute leukemias fuses the ALL-1 gene, related to Drosophila trithorax, to the AF-4 gene.

The ALL-1 gene located at human chromosome 11 band q23 is rearranged in acute leukemias with interstitial deletions or reciprocal translocations between this region and chromosomes 1, 4, 6, 9, 10, or 19. The gene spans approximately 100 kb of DNA and contains at least 21 exons. It encodes a protein of more than 3910 amino acids containing three regions with homology to sequences within the Drosophila trithorax gene, including cysteine-rich regions that can be folded into six zinc finger-like domains. The breakpoint cluster region within ALL-1 spans 8 kb and encompasses several small exons, most of which begin in the same phase of the open reading frame. The t(4;11) chromosome translocation results in two reciprocal fusion products coding for chimeric proteins derived from ALL-1 and from a gene on chromosome 4. This suggests that each 11q23 abnormality gives rise to a specific oncogenic fusion protein.

Amino Acid Sequence↗
PubMed1992

Expression of the normal p53 gene induces differentiation of K562 cells.

The multistep nature of human cancers is well illustrated by chronic myelogenous leukemia (CML), a clonal hematologic malignancy with two distinct phases: chronic and acute. Transition between these phases is characterized by unregulated growth and loss of differentiation of myeloid cells and their progenitors. We recently reported that loss of normal p53 expression correlates with transition from the chronic to acute phase in at least 25% of cases of CML. However, the precise relationship between this loss and biologic features of acute-phase CML is uncertain. To study this question, we artificially expressed normal p53 in K562, an erythroid acute-phase CML cell line lacking normal p53 expression. Biological effects were assessed by determining several growth parameters and by measuring synthesis of hemoglobin, a feature of mature erythroid cells. K562 cells expressing normal p53 had an increased proportion of cells in G1 versus S + G2, a longer doubling time and a lower growth saturation density than control K562 cells or K562 cells with antisense p53. Cells with normal p53 also expressed up to 50-fold more hemoglobin than controls. These data are consistent with the notion that loss of p53 expression may be responsible for many of the features of acute-phase CML cells. The data also demonstrate direct involvement of p53 in differentiation processes.

Cell Cycle↗
PubMed1992

Initiation and progression of chronic myelogenous leukemia.

Different aspects related to initiation of chronic myelogenous leukemia by the t(9;22) translocation and progression of the disease were investigated. Computer search indicated that the repeat within BCR exon I has significant sequence homology to the long terminal repeats of three retroviruses, to two transposons and to the Alu family. This raises the possibility that the BCR repeat is involved in the t(9;22) as well as in generation of the BCR-related loci. Possible involvement of the p53 gene in clinical transition to acute phase was studied. In six patients and cell lines where one allele of the gene was deleted, the other allele was inactivated by loss of transcription, point mutation or rearrangement. The majority of patients, however, have both p53 alleles; detailed analysis of the p53 gene in several of them indicated normal transcription and amino acid sequence.

Base Sequence↗
PubMed1991

p53 in chronic myelogenous leukemia in acute phase.

All patients with chronic myelogenous leukemia (CML) undergo clinical transition from chronic to acute phase. This transition is often associated with deletion of the short arm of chromosome 17 in the form of the i(17q) aberration. Since the p53 gene is a suppressor gene and is located on 17p13, we examined the possibility that it is inactivated during progression of CML. Therefore, we studied the structure and expression of p53 in the leukemic cells of a large number of CML patients in acute phase. We found that although the gene is rarely rearranged, one p53 allele is completely deleted in patients with the i(17q) aberration as well as in some patients who do not show karyotypic changes. In all of these patients the remaining allele is inactivated through loss of expression, rearrangement, or point mutation. Detailed analysis of some patients who carry both p53 alleles indicated neither loss of expression nor structural alterations. It appears that p53 loss of function is associated with progression of around 25% of CML patients.

Base Composition↗
PubMed1989

Analysis of BCR-ABL mRNA in chronic myelogenous leukemia patients and identification of a new BCR-related sequence in human DNA.

The Philadelphia chromosome is present in more than 95% of chronic myelogenous leukemia patients and in up to 25% of patients with acute lymphocytic leukemia. The major consequence of the aberration is the fusion of the ABL and BCR genes. The position of the breakpoint on chromosome 22 determines which species of the potential three fused mRNAs and proteins will be synthesized. We have used the polymerase chain reaction (PCR) to detect these mRNAs in 53 patients and cell lines and found that around 20% contain simultaneously two BCR-ABL mRNAs, presumably due to a process of alternative splicing. The results also indicate that most patients in lymphocytic blast crisis of CML contain the mRNA in which bcr exon 2 is linked to ABL exon II. Finally, we identified, cloned, and characterized a BCR-related sequence that originated from mRNA.

Base Sequence↗
PubMed1989

Nucleotide sequence analysis of human abl and bcr-abl cDNAs.

The complete nucleotide sequence of human abl RNA containing exon Ia was determined. It spans 5598 nucleotides and codes for a protein of 1130 amino acids. The 3' untranslated region contains two short open reading frames and multiple ATTT(A) motifs characteristic of short lived mRNAs. Computer analysis of the abl protein predicts four domains distinct with regard to surface probability and chain flexibility. Nucleotide analysis of the abl segment within a bcr-abl cDNA cloned from the K562 cell line indicated no further alterations within the coding region. A bcr-abl construct containing this segment transformed, together with c-myc, RAT-1 cells and produced a highly active tyrosine kinase.

Amino Acid Sequence↗
PubMed1988

Linear order of the four BCR-related loci in 22q11.

It has recently been shown the a probe for the 3' end of the BCR gene recognizes a family of four BCR-like genes that map to 22q11. Using a panel of somatic cell hybrids with rearrangement of chromosome 22, we have determined their order within 22q11: BCR-2, BCR4, BCR1, BCR-3, with BCR-2 the most centromere proximal. All of the BCR-like genes map proximal to the 22q11-q12 breakpoint of a t(11;22) in a Ewing sarcoma.

Blotting, Southern↗
PubMed1988

Oncogenes in human leukemias.

Eukaryotic cells contain a family of genes termed cellular oncogenes or proto-oncogenes thought to regulate normal cell growth and development. In some abnormal circumstances, such as following transduction by retroviruses, activation of these genes causes leukemias in animals. Possible mechanisms of activation of cellular oncogenes include: point mutation, deletion, or insertion; amplification; activation by internal rearrangement, chromosomal translocation, or promoter insertion; recombinatorial events resulting in the formation of novel chimeric genes; among others. In this review, we consider data implicating activation of cellular oncogenes in the pathogenesis of leukemia in humans. We discuss possible mechanisms whereby oncogene activation may induce leukemias, as well as potential diagnostic and therapeutic implications.

DNA↗
PubMed1988

Molecular biology of chronic myelogenous leukemia.

In this review we have described molecular consequences of the t(9;22) translocation typical of CML and some cases of ALL. This data indicates an important role for abl and bcr and suggest some common mechanisms of activation of c-abl related tyrosine kinase activity. This data also provides insight into the relationship between Ph1 positive, Ph1 negative CML and Ph1 positive ALL. Although this data answers some questions, they raise others; eg, what is the molecular basis of the t(9;22) translocation? How does increased abl related kinase activity eventuate in CML? Finally, this data reviewed suggests that factors other than abl and bcr must play a role in CML. Definition of these factors will be important in the future.

Humans↗
PubMed1988

bcr genes and transcripts.

Human chronic myelogenous leukemia (CML) is a clonal hematologic disorder. CML is characterized by the t(9:22) chromosome translocation which results in translocation of the oncogene abl from chromosome 9 into the breakpoint cluster region (bcr) gene on chromosome 22. We cloned and characterized the cDNA of the normal bcr gene. The bcr gene codes for a protein of 1271 amino acids. The open reading frame is preceded by a region high in GC. At the 5' of this region we identified several GC motifs which are probably involved in the initiation of bcr transcription. bcr transcripts of 7.0 and 4.5 kb are expressed in all cell types examined. These transcripts share all cDNA sequences analysed, including the 5' untranslated region. The latter as well as 902 or 927 amino acids are included within the CML-specific bcr-abl mRNA transcribed from the chimeric bcr-abl gene on chromosome 22. In addition to the complete bcr gene, the human genome contains 3 bcr-related genes containing the last seven exons of the intact bcr gene. One of these genes was analysed in detail and showed high sequence homology with the latter. The three bcr-related genes were probably derived from the intact gene by subsequent steps of duplication.

Base Sequence↗
PubMed1987

Mapping of four distinct BCR-related loci to chromosome region 22q11: order of BCR loci relative to chronic myelogenous leukemia and acute lymphoblastic leukemia breakpoints.

A probe derived from the 3' region of the BCR gene (breakpoint cluster region gene) detects four distinct loci in the human genome. One of the loci corresponds to the complete BCR gene, whereas the others contain a 3' segment of the gene. After HindIII cleavage of human DNA, these four loci are detected as 23-, 19-, 13-, and 9-kilobase-pair fragments, designated BCR4, BCR3, BCR2, and BCR1, respectively, with BCR1 deriving from the original complete BCR gene. All four BCR loci segregate 100% concordantly with human chromosome 22 in a rodent-human somatic cell hybrid panel and are located at chromosome region 22q11.2 by chromosomal in situ hybridization. The BCR2 and BCR4 loci are amplified in leukemia cell line K562 cells, indicating that they fall within the amplification unit that includes immunoglobulin lambda light chain locus (IGL) and ABL locus on the K562 Philadelphia chromosome (Ph1); additionally, in chronic myelogenous leukemia-derived mouse-human hybrids retaining a Ph1 chromosome in the absence of the 9q+ and normal chromosome 22, BCR2 and BCR4 loci are retained, whereas the 3' region of BCR1 and the BCR3 locus are lost, indicating that BCR3 is distal to BCR1 on chromosome 22. Similarly, in mouse-human hybrids retaining a Ph1 chromosome derived from an acute lymphoblastic leukemia-in the absence of the 9q+ and 22, only BCR2 and BCR4 loci are retained, indicating that the breakpoint in this acute lymphoblastic leukemia, as in chronic myelogenous leukemia, is proximal to the BCR1 3' region, but distal to the IGLC locus and the BCR2 and BCR4 3' loci. Thus, the order of loci on chromosome 22 is centromere----BCR2, BCR4, and IGL----BCR1----BCR3----SIS, possibly eliminating BCR2 and BCR4 loci as candidate targets for juxtaposition to the ABL gene in the acute lymphoblastic leukemia Ph1 chromosome.

Chromosome Aberrations↗
PubMed1987

Oncogenes in human leukemias.

Eukaryotic cells contain a family of genes termed cellular oncogenes or proto-oncogenes thought to regulate normal cell growth and development. In some abnormal circumstances, such as following transduction by retroviruses, activation of these genes causes tumors and leukemias in animals. Possible mechanisms of activation of cellular oncogenes include: (1) point mutation, deletion, insertion; (2) amplification; (3) activation by internal rearrangement, chromosomal translocation or promoter insertion, and (4) recombinatorial events resulting in the formation of novel chimeric genes, and others. In this review, we consider data implicating activation of cellular oncogenes in the pathogenesis of leukemia in man. We discuss possible mechanisms whereby oncogene activation may induce leukemias as well as the potential diagnostic and therapeutic implication.

Cell Division↗
PubMed1987

bcr-abl RNA in patients with chronic myelogenous leukemia.

The major consequence of the formation of the Philadelphia (Ph1) chromosome characteristic of leukemia cells of patients with chronic myelogenous leukemia (CML) is fusion of c-abl and bcr genes. Using a sensitive RNase protection technique, we analyzed mRNA from a large number of CML patients. In most, we identified one or both species of bcr-abl chimeric transcripts. These two mRNAs vary in the specific bcr exon joined to abl exon II and are translated into slightly different proteins. The amounts of the fused mRNA within leukemia cells vary considerably between individuals and do not correlate with the phase of the disease.

Exons↗
PubMed1986

Alternative splicing of RNAs transcribed from the human abl gene and from the bcr-abl fused gene.

The primary structure of normal abl protein was determined by sequencing the coding region of its cDNA. abl contains two alternative 5' exons spliced to a common set of 3' exons to yield the two major abl RNA transcripts. These transcripts initiate in different promoter regions and give rise to proteins that vary in their N-termini. In the human cell line K562, abl is translocated from chromosome 9 to within the bcr gene on chromosome 22. Within the fused bcr-abl gene, abl exon II alternatively splices to two adjacent bcr exons. This phenomenon is seen in many patients with chronic myeloid leukemia.

Abelson murine leukemia virus↗
PubMed1979

The transforming gene of Moloney murine sarcoma virus.

A cleavage map of the Moloney murine sarcoma viral DNA was constructed and compared with that of a spontaneously occurring deletion mutant. By restriction enzyme analysis, it was shown that a region encompassing over 40% of the viral information was not essential for transformation or rescue of the deletion mutant. The transforming region was further localised by analysis of the transforming activity in tissue culture of isolated restriction fragments of linear duoble-stranded sarcoma viral DNA. In each case, DNA fragments that retained transforming activity preserved the cell-derived insertion sequences of the viral genome. Moreover, such transformants invariably expressed RNA specific to this region. By these two approaches, it was possible to demonstrate that the transforming region of the viral genome begins very near or within the cell-derived insertion sequences. Thus, the transforming gene of this mammalian sarcoma virus originates from within the mouse cell genome.

Cell Transformation, Viral↗
PubMed1979

Molecular cloning of Moloney murine sarcoma virus: arrangement of virus-related sequences within the normal mouse genome.

The unintegrated circular DNA form of Moloney murine sarcoma virus (MSV) has been cloned in bacteriophage lambda. Discrete deletions in the viral genome were shown to occur during propagation of recombinant phage in Escherichia coli. Heteroduplex and restriction enzyme analyses indicated the deletion of tandemly repeated sequences within certain of the cloned MSV DNA inserts. Cloned MSV DNA was used to prepare a probe composed of its acquired cellular (src) sequences, shown previously to be necessary for MSV transformation. Analysis of EcoRI digests of normal mouse cellular DNA revealed the presence of a single 14-kilobase-pair fragment containing these sequences which lacked contiguity with endogenous type C helper viral information of the same cells. Thus, the sarcoma virus-specific sequences of MSV are represented within the normal mouse genome in a manner analogous to that of a cellular gene.

Animals↗