GENOMICS
6, 192-194
(1990)
SHORT COMMUNICATION The Gene for the Muscle-Specific Enolase Is on the Short Arm of Human Chromosome 17 SALVATORE FEO, *-’ DANIELE OLlvA, t GIOVANNA BARBIERI, t WEIMING MIKE FRIED,* AND AGATA GlALLONGOt
Xu, *
l EukJryotic
Gene Organization and Expression and SSomatic Cell Genetics Laboratories, imperial Cancer Research fund, Lincoln’s Inn Fields, London WC2A 3PX, United Kingdom; and tlstituto di Biologiade/lo Sviluppo, CNR, Via Archirafi, 20, 90123 Palermo, &a/y Received
July 7, 1989;
revised
Enolase (EC 4.2.1.11) catalyzes the interconversion of 2-phosphoglycerate and phosphoenolpyruvate in the glycolytic pathway. In mammals there are at least three distinct forms of enolase that are discriminated by differences in their thermostability, reactivity to various antibodies, electrophoretic mobilities, and tissue distribution (Zomzely-Neurath, 1983). It has been proposed the human enolases are members of a multigene enzyme family (Chen and Giblett, 1976; Pearce et aZ., 1976). By isoenzyme analysis the gene locus ENOl, which encodes the human a- or nonneuronal enolase (NNE), has been mapped to the short arm of chromosome 1 (Khan et al., 1974) and EN02, encoding the human y- or neuron-specific enolase (NSE), was located on chromosome 12 (Grzeschik, 1974; Law and Kao, 1982). Reports of the existence of a third locus, EN03, encoding the /3-enolase, whose gene product is located primarily in adult skeletal muscle, have been based on biochemical and immunological data (Rider and Taylor, 1974). Only recently has the isolation of a cDNA for rat /3-enolasebeen reported (Ohshima et aZ., 1989).
o&w7543/90 Copyright All rights
from Istituto
$3.00 0 1990 by Academic Press, of reproduction in any form
di Biologia
dello
Sviluppo,CNR,
192 Inc. reserved.
14, 1989
While cDNAs for both a- and y-enolase mRNAs have been isolated from different species, the rat gene coding for y-enolase is the only gene that has been cloned and well characterized (Sakimura et aZ., 1987). In our attempts to identify and study the members of the human enolase gene family, we have isolated a number of overlapping genomic clones using (Y-and ycDNAs as probes (Giallongo et aZ., 1986; Oliva et al., 1989). A detailed molecular analysis of the structure, sequence, and expression of one of these clones, which we named XB.21 (Giallongo et aZ., manuscript in preparation), has confirmed that it represents the human gene for ,&enolase. Here we report the chromosomal localization of this gene. A 700-bp XbaI repeat-free fragment (X700), containing the last three exons of the gene, was isolated from the XB.21 phage clone and used as “mapping probe” in this study. On Southern blots of EcoRI-digested human genomic DNA, X700 detected only a 19kb restriction fragment. No cross-reaction with any sequence derived from the other enolase genes was detected under the hybridizing and washing conditions used (data not shown). To determine the chromosomal localization of the human P-enolase gene, the X700 fragment was used to screen Southern blots of EcoRI digested DNA from 12 rodent-human hybrids which contain various complements of human chromosomes. About 15 pg of DNA per lane was separated electrophoretically in 0.8% agarose gels and transblotted to Hybond-N nylon membranes (Amersham):The blots were hybridized at 65°C in a mixture containing 6X SSC (1X SSC is 150 mM NaCl and 15 mM sodium citrate, pH 7.0), 5~ Denhardt’s solution, 1% SDS (sodium dodecyl sulfate), and 2-5 X lo6 cpm/ml of 32P-oligolabeled probe (Feinberg and Vogelstein, 1983) at a specific activity of l-2 X 10’
The human gene encoding the muscle-specific @enolase has been isolated. The &enolase gene was mapped to chromosome 17 by analysis of a panel of rodent-human somatic cell hybrids. The gene was further localized to the short arm and tentatively to the region 17pter-pl l by analysis of cell hybrids and transfectant cell lines carrying different portions of chromosome 17. o 1990 Academic press, I~C.
’ On leave of absence Palermo, Italy.
September
SHORT
cpm/pg. After hybridization the blots were washed for 10 min at room temperature in 2X SSC plus 0.1% SDS, two times for 30 min at 65’C in 2X SSC plus 0.1% SDS, and one time for 30 min at 65°C in 0.2X SSC and then exposed to X-ray film with intensifying screens at -80°C for l-2 days. As shown in Table 1 no discordancies were found for the presence of XB.21 genomic sequence on chromosome 17 and at least three discordancies were found for its presence on any other chromosome; therefore the P-enolase gene can be unambiguously assigned to chromosome 17. This was further confirmed from the hybridization of the X700 probe to DNA from the somatic cell hybrid PCTBA1.8 (Bai et al., 1982) containing only a chromosome 17 as its human counterpart (Fig. 1, lane 1). To further localize the /3-enolase gene on chromosome 17, a second panel of somatic cell hybrids and chromosome-mediated gene transfer transfectants, containing various portions of human chromosome 17 (Xu et al., 1988), were screened with the X700 probe.
TABLE
1
Summary of the Southern Blot Analysis of the DNA of 12 Rodent-Human Somatic Cell Hybrids with the X700 Probe X700
Chromosome
sequence/chromosome retention
+/+
-/Number
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X
3 3 4 3 2 3 3 3 0 3 5 5 3 6 3 2 7 6 0 4 5 4 6
-/+ of hybrid
3 4 0 3 4 2 3 4 5 4 2 4 2 0 5 3 5 2 4 5 4 3 0
+/-
% Discordant
clones 2 1 5 2 1 3 2 1 0 1 3 1 3 5 0 2 0 2 1 0 1 2 5
4 4 3 4 5 4 4 4 7 4 2 2 4 1 4 5 0 1 7 3 2 3 1
193
COMMUNICATION
50.0 41.7 66.7 50.0 50.0 58.3 50.0 41.7 58.3 41.7 41.7 25.0 58.3 50.0 33.3 58.3 0 25.0 66.7 25.0 25.0 41.7 50.0
Note. Hybrids used were CTP412E3, CTP34B4 (9); MOG34A4 (22); 3W4CI5 (8); TwinlQDl2 (16); DT1.2 (23); DUR4.3 (21); HORL411BGP, HORL411B6B4 (1); SIR74ii (25); SIF4A31 (4); PLT1.S (D. Sheer, personal communication).
12345
II
3
4
5
I
I
’
I
I
FIG. 1. Southern blot analysis and schematic representation of the presence or absence of portions of chromosome 17 in the panel of transfectants and somatic cell hybrids used. DNAs were digested with EcoRI, and hybridization was carried out as described in the text. Lane 1: PCTBA1.8 is a mouse-human somatic cell hybrid containing a chromosome 17 as its only human component (2). Lane 2: PJT2/Al is a hybrid containing the APL translocation chromosome (15q+) with the region of 17 below the breakpoint at 17q11.2-q12 (20). Lanes 3 and 4: KLT-8 and PLT-GB, respectively, are transfectant cell lines containing different portions of chromosome 17 (26). Lane 5: TRI-D62 is a mouse-human somatic cell hybrid containing as its only human component the long arm of chromosome 17 (24). Lane 5 was exposed four times longer than the other lanes.
As shown in Fig. 1 only DNA from the cell line PLT6B (lane 4) containing most of the short arm of chromosome 17 hybridized to the 8-enolase-specific probe, positioning the gene in the pter-pll region. The human sarcomeric myosin heavy-chain gene cluster has been mapped to the pter-pll region of chromosome 17 (Leinward et al., 1983; Edwards et al., 1985), while another cluster of myosin heavy-chain genes, specifically expressed in the heart, has been mapped on human chromosome 14 (Saez et al., 1987). It will be of particular interest to know whether the /3enolase gene and the sarcomeric myosin heavy-chain genesare closely linked and whether their chromosomal position has a role in the tissue-specific regulation of these genes. ACKNOWLEDGMENTS We thank N. Spurr for providing us with the somatic cell hybrid DNAs and E. Solomon for her advice and useful comments during the preparation of the manuscript. This work was supported partially by Grant 113/M from Progetti di Ricerca Sanitaria Finalizzata, Regione Siciliana, to A.G.
REFERENCES 1.
2.
ANDREW& P. W., KNOWLES, B. B., PARKAR, M., PYM, B., STANLEY, K., AND GOODFELLOW, P. N. (1985). A human cellsurface antigen defined by monoclonal antibodies and controlled by a gene on human chromosome 1. Ann. Hum. Genet. 49: 3139. BAI, Y., SHEER, D., HIORNS, L., KNOWLES, R. W., AND TUNNACLIFFE, A. (1982). A monoclonal antibody recognizing a cell
194
SHORT
COMMUNICATION
surface antigen coded by a gene on human chromosome 17. Ann.
Hum.
Genet.
46: 337-347.
3. CHEN, S., AND GIBLETT, E. R. (1976). Enolase: Human tissue distribution and evidence for three different loci. Ann. Hum. Genet.
39:
277-280.
4. EDWARDS, Y. H., PARKAR, M., POVEY, S., WEST, L. F., PARRINGTON,J. M., AND SOLOMON, E. (1985). Human myosin heavy chain genes assigned to chromosome 17 using a human cDNA as probe. Ann. Hum. Genet. 49: 101-109. 5. FEINBERG, A. P., AND VOGELSTEIN, B. (1983). A technique for radiolabeling restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6-13. 6. GIALLONGO, A., FEo, S., MOORE, R., CROCE,C. M., AND SHOWE, L. C. (1986). Molecular cloning and nucleotide sequence of a full-length cDNA for human alpha enolase. Proc. Natl. Acad. Sci. USA
83: 6741-6745.
7. GRZESCHIK,K. H. (1974). Assignment of human genes: @-Glucuronidase to chromosome 7, adenylate kinase-1 to 9, a second enzyme with enolase activity to 12, and mitochondrial IDH to 15. Cytogenet. Cell Genet. 16: 142-148. 8. HOBART, M. J., RAF%BITTS,T. H., GOODFELLOW, P. N., SOLOMON, E., CHAMBERS, S., SPURR, N., AND POVEY, S. (1981). Immunoglobulin heavy chain genes in human are located on chromosome 14. Ann. Hum. Genet. 46: 331-335. 9. JONES, E. A., GOODFELLOW, P. N., KENNETT, R. H., AND BODMER, W. F. (1976). The independent expression of HLA and /32-microglobulin on human-mouse hybrids. Somat. Cell Genet. 2:483-496. 10. KHAN, M., DOOPERT, B. A., HAGMMEIJER, A., AND WESTERVELD, A. (1974). The human loci for phosphopyruvate hydratase and guanylate kinase are syntenic with the PGD-PGM, linkage group in man-Chinese hamster somatic cell hybrids. Cytogenet. Cell Genet. 13: 130-136. 11. LAW, M. L., AND KAO, F. (1982). Regional mapping of the gene coding for enolase-2 on human chromosome 12. J. Cell Sci. 53: 245-254. 12. LEINWARD, L. A., FOURNIER,R. E. K., NADAL-GINARD, B., AND SHOWS, T. B. (1983). Multigene family for sarcomeric myosin heavy chain in mouse and human: Localization on a single chromosome. Science 221: 766-769. 13. OHSHIMA, Y., MITSUI, H., TAKAYAMA, Y., KUSHIYA, E., SAKIMURA, K., AND TAKAHASHI, Y. (1989). cDNA cloning and nucleotide sequence of rat muscle specific enolase (j3p enolase). FEBS
Lett.
2: 425-430.
14. OLIVA, D., BARBA, G., BARBIERI, G., GIALLONGO, A., AND FEO, S. (1989). Cloning, expression and sequence homologies of cDNA for human gamma enolase. Gene 79: 355-360. 15. PEARCE,J. M., EDWARDS,Y. H., AND HARRIS, H. (1976). Human enolase isoenzymes: Electrophoretic and biochemical evidence for three loci. Ann. Hum. Genet. 39: 263-276.
16. PHILLIPS, I. R., SHEPHARD, E. A., POVEY, S., DAVIES, M. B., KELSEY, G., MANTEIRO, M., WEST, L. F., AND COWELL, F. (1985). A cytochrome P-450 gene family mapped to human chromosome 19. Ann. Hum. Genet. 49: 267-274. 17. RIDER, C. C., AND TAYLOR, C. B. (1974). Enolase isoenzymes in rat tissue: Electrophoretic, chromatographic, immunological and kinetic properties. Biochem. Biophys. Acta 365: 285-300. 18. SAEZ, L. J., GIANOLA, R. M., MCNELLY, E. M., FEGHALI, R., EDDY, R., SHOWS, T. B., AND LEINWARD, L. A. (1987). Human cardiac myosin heavy chain genes and their linkage in the genome. Nucleic Acids Res. 15: 5443-5449. 19. SAKIMURA, K., KUSHIYA, E., TAKAHASHI, Y., AND SUZUKI, Y. (1987). The structure and expression of neuron-specific enolase gene. Gene 60: 103-113. 20. SHEER, D., HIORNS, L. R., STANLEY, K. F., GOODFELLOW, P. N., SWALLOW, D. M., POWEY, S., HEISTERKAMP, N., GROFFEN, J., STEPHENSON, J. R., AND SOLOMON, E. (1983). Genetic analysis of the 15;17 chromosome translocation associated with acute promyelocytic leukemia. Proc. N&l. Acad. Sci. USA 80: 5007-5011. 21. SOLOMON, E., BOROW, M., GOODFELLOW, P. N., BODMER, W. F., SWALLOW, D. M., POVEY, S., AND NOEL, B. (1976). Human gene mapping using an X/autosome translocation. Some. Cell Genet. 2: 125-140. 22. SOLOMON, E., SWALLOW, D., BURGESS, S., AND EVANS, L. (1979). Assignment of the human acid cu-glucosidase gene ((YGLU) to chromosome 17 using somatic cell hybrids. Ann. Hum. Genet.
42:
273-281.
23. SWALLOW, D. M., SOLOMON, E., AND PAJUNEN, L. (1977). Immunochemical analysis of the N-acetyl hexosaminidaaes in human-mouse hybrids made using a double selective system. Cytogenet. Cell Genet. 18: 136-148. 24. TUNNACLIFFE, A., PARKAR, M., POVEY, S., BENGTSSON, B. O., STANLEY, K., SOLOMON, E., AND GOODFELLOW, P. N. (1983). Integration of Ecogpt and SV40 early region sequences into human chromosome 17: A dominant selection system in whole cells and microcell human-mouse hybrids. EMBO J. 2: 15771584. 25. WHITEHEAD, A. S., SOLOMON, E., CHAMBERS, S., BODMER, W. F., POVEY, S., AND FEY, G. (1982). Assignment of the structural gene for the third component of human complement to chromosome 19. Proc. Natl. Acad, Sci. USA 79: 5021-5025. 26. Xv, W., GORMAN, P. A., RIDER, S. H., HEDGE, P. J., MOORE, G., PRICHARDS,C., SHEER, D., AND SOLOMON, E. (1988). Construction of a genetic map of human chromosome 17 by use of chromosome-mediated gene transfer. Proc. Nat1 Acad. Sci. USA 85:8563-8567. 27. ZOMZELY-NEURATH, C. E. (1983). Enolase. In “Handbook of Neurochemistry” (A. Lajtha, Ed.), Vol. 4,2nd ed., pp. 403-433, Plenum, New York.
6, 192-194
(1990)
SHORT COMMUNICATION The Gene for the Muscle-Specific Enolase Is on the Short Arm of Human Chromosome 17 SALVATORE FEO, *-’ DANIELE OLlvA, t GIOVANNA BARBIERI, t WEIMING MIKE FRIED,* AND AGATA GlALLONGOt
Xu, *
l EukJryotic
Gene Organization and Expression and SSomatic Cell Genetics Laboratories, imperial Cancer Research fund, Lincoln’s Inn Fields, London WC2A 3PX, United Kingdom; and tlstituto di Biologiade/lo Sviluppo, CNR, Via Archirafi, 20, 90123 Palermo, &a/y Received
July 7, 1989;
revised
Enolase (EC 4.2.1.11) catalyzes the interconversion of 2-phosphoglycerate and phosphoenolpyruvate in the glycolytic pathway. In mammals there are at least three distinct forms of enolase that are discriminated by differences in their thermostability, reactivity to various antibodies, electrophoretic mobilities, and tissue distribution (Zomzely-Neurath, 1983). It has been proposed the human enolases are members of a multigene enzyme family (Chen and Giblett, 1976; Pearce et aZ., 1976). By isoenzyme analysis the gene locus ENOl, which encodes the human a- or nonneuronal enolase (NNE), has been mapped to the short arm of chromosome 1 (Khan et al., 1974) and EN02, encoding the human y- or neuron-specific enolase (NSE), was located on chromosome 12 (Grzeschik, 1974; Law and Kao, 1982). Reports of the existence of a third locus, EN03, encoding the /3-enolase, whose gene product is located primarily in adult skeletal muscle, have been based on biochemical and immunological data (Rider and Taylor, 1974). Only recently has the isolation of a cDNA for rat /3-enolasebeen reported (Ohshima et aZ., 1989).
o&w7543/90 Copyright All rights
from Istituto
$3.00 0 1990 by Academic Press, of reproduction in any form
di Biologia
dello
Sviluppo,CNR,
192 Inc. reserved.
14, 1989
While cDNAs for both a- and y-enolase mRNAs have been isolated from different species, the rat gene coding for y-enolase is the only gene that has been cloned and well characterized (Sakimura et aZ., 1987). In our attempts to identify and study the members of the human enolase gene family, we have isolated a number of overlapping genomic clones using (Y-and ycDNAs as probes (Giallongo et aZ., 1986; Oliva et al., 1989). A detailed molecular analysis of the structure, sequence, and expression of one of these clones, which we named XB.21 (Giallongo et aZ., manuscript in preparation), has confirmed that it represents the human gene for ,&enolase. Here we report the chromosomal localization of this gene. A 700-bp XbaI repeat-free fragment (X700), containing the last three exons of the gene, was isolated from the XB.21 phage clone and used as “mapping probe” in this study. On Southern blots of EcoRI-digested human genomic DNA, X700 detected only a 19kb restriction fragment. No cross-reaction with any sequence derived from the other enolase genes was detected under the hybridizing and washing conditions used (data not shown). To determine the chromosomal localization of the human P-enolase gene, the X700 fragment was used to screen Southern blots of EcoRI digested DNA from 12 rodent-human hybrids which contain various complements of human chromosomes. About 15 pg of DNA per lane was separated electrophoretically in 0.8% agarose gels and transblotted to Hybond-N nylon membranes (Amersham):The blots were hybridized at 65°C in a mixture containing 6X SSC (1X SSC is 150 mM NaCl and 15 mM sodium citrate, pH 7.0), 5~ Denhardt’s solution, 1% SDS (sodium dodecyl sulfate), and 2-5 X lo6 cpm/ml of 32P-oligolabeled probe (Feinberg and Vogelstein, 1983) at a specific activity of l-2 X 10’
The human gene encoding the muscle-specific @enolase has been isolated. The &enolase gene was mapped to chromosome 17 by analysis of a panel of rodent-human somatic cell hybrids. The gene was further localized to the short arm and tentatively to the region 17pter-pl l by analysis of cell hybrids and transfectant cell lines carrying different portions of chromosome 17. o 1990 Academic press, I~C.
’ On leave of absence Palermo, Italy.
September
SHORT
cpm/pg. After hybridization the blots were washed for 10 min at room temperature in 2X SSC plus 0.1% SDS, two times for 30 min at 65’C in 2X SSC plus 0.1% SDS, and one time for 30 min at 65°C in 0.2X SSC and then exposed to X-ray film with intensifying screens at -80°C for l-2 days. As shown in Table 1 no discordancies were found for the presence of XB.21 genomic sequence on chromosome 17 and at least three discordancies were found for its presence on any other chromosome; therefore the P-enolase gene can be unambiguously assigned to chromosome 17. This was further confirmed from the hybridization of the X700 probe to DNA from the somatic cell hybrid PCTBA1.8 (Bai et al., 1982) containing only a chromosome 17 as its human counterpart (Fig. 1, lane 1). To further localize the /3-enolase gene on chromosome 17, a second panel of somatic cell hybrids and chromosome-mediated gene transfer transfectants, containing various portions of human chromosome 17 (Xu et al., 1988), were screened with the X700 probe.
TABLE
1
Summary of the Southern Blot Analysis of the DNA of 12 Rodent-Human Somatic Cell Hybrids with the X700 Probe X700
Chromosome
sequence/chromosome retention
+/+
-/Number
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X
3 3 4 3 2 3 3 3 0 3 5 5 3 6 3 2 7 6 0 4 5 4 6
-/+ of hybrid
3 4 0 3 4 2 3 4 5 4 2 4 2 0 5 3 5 2 4 5 4 3 0
+/-
% Discordant
clones 2 1 5 2 1 3 2 1 0 1 3 1 3 5 0 2 0 2 1 0 1 2 5
4 4 3 4 5 4 4 4 7 4 2 2 4 1 4 5 0 1 7 3 2 3 1
193
COMMUNICATION
50.0 41.7 66.7 50.0 50.0 58.3 50.0 41.7 58.3 41.7 41.7 25.0 58.3 50.0 33.3 58.3 0 25.0 66.7 25.0 25.0 41.7 50.0
Note. Hybrids used were CTP412E3, CTP34B4 (9); MOG34A4 (22); 3W4CI5 (8); TwinlQDl2 (16); DT1.2 (23); DUR4.3 (21); HORL411BGP, HORL411B6B4 (1); SIR74ii (25); SIF4A31 (4); PLT1.S (D. Sheer, personal communication).
12345
II
3
4
5
I
I
’
I
I
FIG. 1. Southern blot analysis and schematic representation of the presence or absence of portions of chromosome 17 in the panel of transfectants and somatic cell hybrids used. DNAs were digested with EcoRI, and hybridization was carried out as described in the text. Lane 1: PCTBA1.8 is a mouse-human somatic cell hybrid containing a chromosome 17 as its only human component (2). Lane 2: PJT2/Al is a hybrid containing the APL translocation chromosome (15q+) with the region of 17 below the breakpoint at 17q11.2-q12 (20). Lanes 3 and 4: KLT-8 and PLT-GB, respectively, are transfectant cell lines containing different portions of chromosome 17 (26). Lane 5: TRI-D62 is a mouse-human somatic cell hybrid containing as its only human component the long arm of chromosome 17 (24). Lane 5 was exposed four times longer than the other lanes.
As shown in Fig. 1 only DNA from the cell line PLT6B (lane 4) containing most of the short arm of chromosome 17 hybridized to the 8-enolase-specific probe, positioning the gene in the pter-pll region. The human sarcomeric myosin heavy-chain gene cluster has been mapped to the pter-pll region of chromosome 17 (Leinward et al., 1983; Edwards et al., 1985), while another cluster of myosin heavy-chain genes, specifically expressed in the heart, has been mapped on human chromosome 14 (Saez et al., 1987). It will be of particular interest to know whether the /3enolase gene and the sarcomeric myosin heavy-chain genesare closely linked and whether their chromosomal position has a role in the tissue-specific regulation of these genes. ACKNOWLEDGMENTS We thank N. Spurr for providing us with the somatic cell hybrid DNAs and E. Solomon for her advice and useful comments during the preparation of the manuscript. This work was supported partially by Grant 113/M from Progetti di Ricerca Sanitaria Finalizzata, Regione Siciliana, to A.G.
REFERENCES 1.
2.
ANDREW& P. W., KNOWLES, B. B., PARKAR, M., PYM, B., STANLEY, K., AND GOODFELLOW, P. N. (1985). A human cellsurface antigen defined by monoclonal antibodies and controlled by a gene on human chromosome 1. Ann. Hum. Genet. 49: 3139. BAI, Y., SHEER, D., HIORNS, L., KNOWLES, R. W., AND TUNNACLIFFE, A. (1982). A monoclonal antibody recognizing a cell
194
SHORT
COMMUNICATION
surface antigen coded by a gene on human chromosome 17. Ann.
Hum.
Genet.
46: 337-347.
3. CHEN, S., AND GIBLETT, E. R. (1976). Enolase: Human tissue distribution and evidence for three different loci. Ann. Hum. Genet.
39:
277-280.
4. EDWARDS, Y. H., PARKAR, M., POVEY, S., WEST, L. F., PARRINGTON,J. M., AND SOLOMON, E. (1985). Human myosin heavy chain genes assigned to chromosome 17 using a human cDNA as probe. Ann. Hum. Genet. 49: 101-109. 5. FEINBERG, A. P., AND VOGELSTEIN, B. (1983). A technique for radiolabeling restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6-13. 6. GIALLONGO, A., FEo, S., MOORE, R., CROCE,C. M., AND SHOWE, L. C. (1986). Molecular cloning and nucleotide sequence of a full-length cDNA for human alpha enolase. Proc. Natl. Acad. Sci. USA
83: 6741-6745.
7. GRZESCHIK,K. H. (1974). Assignment of human genes: @-Glucuronidase to chromosome 7, adenylate kinase-1 to 9, a second enzyme with enolase activity to 12, and mitochondrial IDH to 15. Cytogenet. Cell Genet. 16: 142-148. 8. HOBART, M. J., RAF%BITTS,T. H., GOODFELLOW, P. N., SOLOMON, E., CHAMBERS, S., SPURR, N., AND POVEY, S. (1981). Immunoglobulin heavy chain genes in human are located on chromosome 14. Ann. Hum. Genet. 46: 331-335. 9. JONES, E. A., GOODFELLOW, P. N., KENNETT, R. H., AND BODMER, W. F. (1976). The independent expression of HLA and /32-microglobulin on human-mouse hybrids. Somat. Cell Genet. 2:483-496. 10. KHAN, M., DOOPERT, B. A., HAGMMEIJER, A., AND WESTERVELD, A. (1974). The human loci for phosphopyruvate hydratase and guanylate kinase are syntenic with the PGD-PGM, linkage group in man-Chinese hamster somatic cell hybrids. Cytogenet. Cell Genet. 13: 130-136. 11. LAW, M. L., AND KAO, F. (1982). Regional mapping of the gene coding for enolase-2 on human chromosome 12. J. Cell Sci. 53: 245-254. 12. LEINWARD, L. A., FOURNIER,R. E. K., NADAL-GINARD, B., AND SHOWS, T. B. (1983). Multigene family for sarcomeric myosin heavy chain in mouse and human: Localization on a single chromosome. Science 221: 766-769. 13. OHSHIMA, Y., MITSUI, H., TAKAYAMA, Y., KUSHIYA, E., SAKIMURA, K., AND TAKAHASHI, Y. (1989). cDNA cloning and nucleotide sequence of rat muscle specific enolase (j3p enolase). FEBS
Lett.
2: 425-430.
14. OLIVA, D., BARBA, G., BARBIERI, G., GIALLONGO, A., AND FEO, S. (1989). Cloning, expression and sequence homologies of cDNA for human gamma enolase. Gene 79: 355-360. 15. PEARCE,J. M., EDWARDS,Y. H., AND HARRIS, H. (1976). Human enolase isoenzymes: Electrophoretic and biochemical evidence for three loci. Ann. Hum. Genet. 39: 263-276.
16. PHILLIPS, I. R., SHEPHARD, E. A., POVEY, S., DAVIES, M. B., KELSEY, G., MANTEIRO, M., WEST, L. F., AND COWELL, F. (1985). A cytochrome P-450 gene family mapped to human chromosome 19. Ann. Hum. Genet. 49: 267-274. 17. RIDER, C. C., AND TAYLOR, C. B. (1974). Enolase isoenzymes in rat tissue: Electrophoretic, chromatographic, immunological and kinetic properties. Biochem. Biophys. Acta 365: 285-300. 18. SAEZ, L. J., GIANOLA, R. M., MCNELLY, E. M., FEGHALI, R., EDDY, R., SHOWS, T. B., AND LEINWARD, L. A. (1987). Human cardiac myosin heavy chain genes and their linkage in the genome. Nucleic Acids Res. 15: 5443-5449. 19. SAKIMURA, K., KUSHIYA, E., TAKAHASHI, Y., AND SUZUKI, Y. (1987). The structure and expression of neuron-specific enolase gene. Gene 60: 103-113. 20. SHEER, D., HIORNS, L. R., STANLEY, K. F., GOODFELLOW, P. N., SWALLOW, D. M., POWEY, S., HEISTERKAMP, N., GROFFEN, J., STEPHENSON, J. R., AND SOLOMON, E. (1983). Genetic analysis of the 15;17 chromosome translocation associated with acute promyelocytic leukemia. Proc. N&l. Acad. Sci. USA 80: 5007-5011. 21. SOLOMON, E., BOROW, M., GOODFELLOW, P. N., BODMER, W. F., SWALLOW, D. M., POVEY, S., AND NOEL, B. (1976). Human gene mapping using an X/autosome translocation. Some. Cell Genet. 2: 125-140. 22. SOLOMON, E., SWALLOW, D., BURGESS, S., AND EVANS, L. (1979). Assignment of the human acid cu-glucosidase gene ((YGLU) to chromosome 17 using somatic cell hybrids. Ann. Hum. Genet.
42:
273-281.
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