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Subregional Localization of the Gene(s) Governing the Human
Interferon Induced Antiviral State in Man
SUMMARY
A dosage effect of chromosomal translocation was used to locate the gene(s)
which codes for the human interferon induced antiviral state on the long arm of
chromosome 2I.
Using mouse-human somatic hybrid cells, Tan, Tischfield & Ruddle (1973) assigned
the gene(s) which codes for the human interferon (HIF) induced antiviral state (AVS)
to chromosome 2I. Presently, it is not clear if the product of this gene assignment is the
putative antiviral protein, the putative receptor site(s) for HIF, or both (Chany et al. J975;
Tan, I975). Nevertheless, the assignment has been confirmed by the dosage effect of chromosome 21 on the inducibility of the AVS in human cells (Tan et al. 1974; Tan, I975). In this
paper we report localization of the gene(s) in question to the long arm of chromosome 21 by
measuring the dosage effect of translocated chromosome 21 on the inducibility of the AVS
in human cells.
Four human skin fibroblast lines with translocated chromosomes were used. (i) A diploid
fibroblast line (t4-2I) in which one of its two chromosomes 21 is translocated to the short
arm of chromosome 4 (Seabright et al. 1975). The translocation results in the deletion of the
short arm of chromosome 2~. (ii) A trisomic 2I fibroblast line (t6-zx) in which one of its
three chromosomes 21 is translocated to the short arm of chromosome 6. The translocation
also results in the deletion of the short arm of chromosome 2I (Borganker, Greene & Coriell,
I974). (iii) A diploid cell line (t2-4), kindly supplied by Dr P. Bowen, University of Alberta,
in which a part of chromosome 2 is translocated to chromosome 4. (iv) A trisomic 2I cell
line (tI-I7) in which chromosome I is translocated to chromosome I7 (Chapelle et al. 1975).
In addition to these lines, eight trisomic 2 I (Y-2I), ten normal diploid (D-21), one monosomic 2I (M-zI) and one trisomic I8 (T-I8) lines of skin fibroblasts were used.
These fibroblast cultures were individually treated with various amounts of human leucocyte interferon. Two preparations were used: (I) one provided by Dr J. Valenta, Smith
Kline & French, and partially purified on a Sephadex G-75 column to asp. act. of approx.
4 × lO5 N I H reference units/rag protein (SKF-HIF), and (2) the N I H leucocyte interferon
reference reagent, code Go23-9o 1-527 (NIH-HIF). The inducibility of the AVS was assessed
in terms of the amount of H I F required to inhibit vesicular stomatitis virus (VSV) R N A
synthesis in these cells by 5o ~ (Tan, 1975). When a given cell line was tested in turn with
each interferon preparation, it was found that some 6 to 8 times more units of the N I H
reagent interferon than of the SKF preparation seemed consistently to be needed for 5o ~o
inhibition of VSV-RNA synthesis (the reasons for this were not investigated, but presumably
reflect assay standardization problems). However, when different cell lines were compared
the relative number of units of each type of interferon preparation required for 50 %oinhibition were closely comparable, as shown in Table I. The results in this~table show that the
t4-21 cells required about the same amount of HIF as D-21 cells whereas t6-21 cells required
about the same amount of H I F as T-2I to induce the AVS. Previously, it has been shown
(Tan, 1975) that the inducibility of the AVS in human cells is related to the number of
chromosomes 2~ present in the cell. According to this finding, one would expect the t4-21
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Table i. Chromosome 2t dosage effect on the inducibility of the
anti-viral state in human fibroblasts
Units of human interferon inhibiting VSV-RNA
synthesis by 50% *
f
Cell type
Identification number
SKF preparation
NIH preparation
M-2I
GM-23o
0"28, 0"26, 0"29
1'6, 1"9
D-2 r
G M-38
GM-I81
GM-72
GM-I79
GM-37
G M-71
G M-638
No. I41
GRC I5
GRC 1I5
o'o62
0"064, o'o59
o'062
O'O58
O'O66
0059
o'o59
0-070, 0"069
o.o66
0"069
o'45
0"50, 0"43
O'49
o'4I
o'49
0"43
o'5o
0"42
N.D.t
o'48, 0"45
t4-21
t2-4
GM-98
GM-3Ol
o'055, o'o59
o.o61, o'o62
0"38, o'39, o'45
o'4o, o'44
T-I8
T-2I
GM-I43
GM-258
GRC t4
GRC It4
DSGG I
DSGG 2
o'o7o, o'o7o
o'o16
o.o18
o'oi4
0"018
0"017
o'48, o'5o
o.Io, o-I I
N.D.
o't2
N.D.
N.D.
N o . 21
0"018
N.D.
No. 23
No. 25
o'o18
o"o16
N.D.
N.D.
t6-Zl
GM-144
O'OLO, o'o12, o'oi2
o'o8, o'o9
T-21(ttqT)
GM-2ol
o'oi4, o'o16
o'o9, o ' I t
* Each figure represents the results from one determination.
t Not done.
and t6-2I cells to require about the same amount of H I F to induce the AVS as M-2! and
D-2I cells, respectively, if the gene(s) for the induction of the AVS is located on the short
arm of chromosome zI. On the contrary, if the gene(s) is on the long arm of chromosome
2I, one would expect the t4-2I and t6-2i cells to require about the same amounts Of H I F as
D-2t and T-z1 cells, respectively. Our results in Table I favour the hypothesis that the gene(s)
for the induction of the AVS is on the long arm of chromosome z~.
The translocated piece of chromosome 2i in the t6-2~ cells was cytogenetically verified
(a routine examination performed at the cell repository at Camden) to consist of most of
the long arm of this chromosome, as previously reported by Borganker et al. 0974). The
fact that this translocated portion contributed to a chromosome 2I dosage effect on the
inducibility of the AVS by H I F in these cells indicates that the gene(s) for AVS is located
within its limits. It is unlikely that the dosage effect is the result of a non-specific translocated
condition, or an addition of a human chromosome other than chromosome 2I because the
T-zl(t~-t7) line and the T-18 line as well as the D-z1(t2-4) line required about the same
amounts of H I F to be induced as the T-21 and D-2I lines, respectively (Table I). The translocated T-21 (t-17) cells have three copies of chromosome 2I, and the T-t8 cells and the t2- 4
cells have two copies of chromosome 2i. In view of these considerations we wish to suggest
that the gene(s) governing the inducibility of the AVS is on the long arm of chromosome 21.
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A gene dosage effect o n a gene p r o d u c t has also been shown between c h r o m o s o m e 16 a n d
adenine p h o s p h o r i b o s y l transferase ( M a r i m o & Gianelli, 1975) as well as between superoxide
dismutase (cytosol form) and c h r o m o s o m e 21 (Sichitiu et aL 1974). U n d o u b t e d l y , c h r o m o s o m e dosage effects on a variety o f o t h e r assigned gene p r o d u c t s will be r e p o r t e d in the near
future. It should then be feasible to consider the use o f t r a n s l o c a t e d c h r o m o s o m a l dosage for
the sub-regional m a p p i n g o f other genetic m a r k e r s on to h u m a n c h r o m o s o m e s . One obvious
l i m i t a t i o n with this technique is the availability o f n a t u r a l l y occurring translocations. However, such limitations can possibly be resolved b y generating a p p r o p r i a t e c h r o m o s o m a l
deletions by the m i c r o b e a m l a s e r - i r r a d i a t i o n technique o f Berns (1974). These artificially
generated deletions, in t u r n c o u l d be used for m a p p i n g studies b y m e a s u r i n g the dosage
effects o f these deletions on a specific gene product.
Division o f Medical Biochemistry
Faculty o f Medicine
University o f Calgary
Calgary, Alberta T2N I N 4
Canada
Y . H . TAN
Institute for Medical Research
Camden, New Jersey, U.S.A.
A . E . GREENE
REFERENCES
BERNS,M. W. (1974)" Directed chromosomal loss by micro-laser irradiation. Science, New York x86, 700-705.
BORGANKER,D. S., GREENE,A. E. & CORIELL,L. L. 0974). A (6:2I) translocation, unbalanced, 46t chromosomes. Cytogenetics and Cell Genetics x3, 4o3-4o5.
CHANY, C., VIGNAL, M., COUILLIN, P., VAN CONG, N., BOUle, J. & BOUle, A. 0975). Chromosomal localization of
human gene(s) governing the interferon induced anti-viral state. Proceedingsof the National Academy of
Sciences of the United States of America 72, 3129-3133
CHAPELLE, A. DE LA, MILLER, R. C., GREENE, A. E. & CORIELL, L. L. (1975). A ( 1 " 1 7 ) translocation, unbalanced,
plus trisomy 2I, 47 chromosomes. Cytogenetics and Cell Genetics I4, 82-83.
MARIMO,B. & GIANELLI,F. (1975). Gene dosage effect in human trisomy I6. Nature, London z56, 204-206.
SEABRIGHT, M., MILLER, R. C., GREENE, A. E. & CORIELL, L. L. (I975). A (4:21) translocation, unbalanced, 46
chromosomes. Cytogenetics and Cell Genetics x4, I52-I53.
SICH1TIU, S., S1NET, P. M., LEJEUNE, J. & FREZAL. (I974). Surdosage de la forme dimerique de l'indophenoloxydase dans la trisomie 2I, secondaire au surdosage g6nique. Humangenetik z3, 65-72.
TAN, V. H. 0975): Dosage effect of chromosome 21 on the inducibility of the anti-viral gene(s). Nature z53,
280-282.
TAN, Y. H., SCHNEIDER, E. L., TISCHFIELD, J. A., EPSTEIN, C. & RUDDLE, F. H. 0974)- D e m o n s t r a t i o n o f c h r o m o s o m e 21 gene dosage in man. Science, New York I8.5, 61-63.
TAN, Y. H., TISCHFIELD, J. A. & RUDDLE, F. H. (1973). The linkage of genes for human interferon induced anti-
viral protein(s) and superoxide dismutase cytosol to chromosome 21. Journal of Experimental Medicine
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(Received 4 November 1975)
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