Adult and fetal human globin genes are expressed following chromosomal transfer into MEL cells.
Somatic cell hybridization of mouse erythroleukemia (MEL) cells and HEL cells, a human erythroleukemia line that produces fetal (gamma) but fails to express adult (beta) globin, was used to test whether the expression of the two human globin genes is regulated cis or trans. An experimental approach using anti-human globin monoclonal antibodies for detection, efficient cloning, and monitoring of hybrids of interest was employed. Further characterization of hybrids used isoelectric focusing for detection of human globins and S1 nuclease mapping. In contrast to the parental HEL line, all chromosome 11-retaining HEL-MEL hybrids expressed human beta-globin, suggesting that the HEL beta-globin genes (i) are transcriptionally competent, (ii) become activated in response to a positive trans-acting element within the MEL environment, and (iii) fail to express into the HEL environment because of either the absence of a positive trans-acting element or the presence of a trans-acting inhibitor of beta-globin gene expression. In addition to beta-globin, the primary HEL-MEL hybrids co-expressed gamma-globin; however, gamma-globin expression segregated by subcloning so that secondary and tertiary clones either expressed only beta-globin or co-expressed gamma- and beta-globin. The results of subcloning can be explained by assuming that gamma-globin gene expression is controlled by a HEL cell-derived transacting element encoded by a gene not syntenic to chromosome 11 or by postulating that the HEL gamma-globin genes become randomly modified during the continuous proliferation of hybrids.
Papayannopoulou
T H
TH
Lindsley
D
D
Kurachi
S
S
Lewison
K
K
Hemenway
T
T
Melis
M
M
Anagnou
N P
NP
Najfeld
V
V
D000818:Animals; D000911:Antibodies, Monoclonal; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D005260:Female; D005319:Fetal Hemoglobin / Q000235:genetics; D005470:Fluorometry; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism*; D007525:Isoelectric Focusing; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D051379:Mice; D011247:Pregnancy; D015719:Single-Strand Specific DNA and RNA Endonucleases
D000911:Antibodies, Monoclonal; D005914:Globins; D005319:Fetal Hemoglobin; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases