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J Virol, August 1998, p. 6898-6901, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Binding of Murine Leukemia Virus Gag Polyproteins
to KIF4, a Microtubule-Based Motor Protein
Wankee
Kim,1,2
Yao
Tang,1
Yasushi
Okada,3
Ted A.
Torrey,1
Sisir K.
Chattopadhyay,1
Michael
Pfleiderer,4
Falko G.
Falkner,4
Friedrich
Dorner,4
Wonja
Choi,5
Nobutaka
Hirokawa,3 and
Herbert C.
Morse III1,*
Laboratory of Immunopathology, National
Institute of Allergy and Infectious Diseases, Bethesda, Maryland
208921;
Department of Cell Biology and
Anatomy, Graduate School, University of Tokyo, Tokyo,
Japan3;
IMMUNO AG, Vienna,
Austria4; and
Molecular Biology
Laboratory, Institute for Medical Sciences, Ajou University,
Kyongki,2 and
Ewha Women's
University, Seoul,5 Korea
Received 29 October 1997/Accepted 12 May 1998
 |
ABSTRACT |
A cDNA clone encoding a cellular protein that interacts with murine
leukemia virus (MuLV) Gag proteins was isolated from a T-cell lymphoma
library. The sequence of the clone is identical to the C terminus of a
cellular protein, KIF4, a microtubule-associated motor protein that
belongs to the kinesin superfamily. KIF4-MuLV Gag associations have
been detected in vitro and in vivo in mammalian cells. We suggest that
KIF4 could be involved in Gag polyprotein translocation from the
cytoplasm to the cell membrane.
| |
TEXT |
The gag gene of murine
leukemia viruses (MuLVs) encodes a Pr65Gag polyprotein that
is responsible for virion particle formation, assembly, and budding at
the cell membrane (reviewed in reference 26).
Although N-terminal myristylation of the polyprotein has been
identified as critical for plasma membrane targeting, many details of
Gag protein folding, transport, membrane binding, and assembly into
MuLV particles are not fully understood (11).
Previously, it was found that an immunodeficiency syndrome of mice,
murine AIDS, is induced by a Pr60 variant of Gag encoded by a
replication-defective virus designated BM5def (2) or Du5H (1). To explore the mechanisms by which the
Pr60Gag protein of BM5def contributes to murine AIDS, we
utilized the yeast two-hybrid system to screen cellular proteins that
are capable of binding to this unique Gag protein. During the course of
this screening, we found that the Pr60Gag protein of BM5def
and the Pr65Gag protein of ecotropic MuLV bind to a
cellular protein called KIF4 (21), a member of the kinesin
superfamily of motor proteins (9, 12-14).
Previous studies have shown that KIF4 is a ubiquitously expressed
protein especially abundant in juvenile neurons and lymphatic tissue
(21). It has a microtubule plus-end-directed motor activity and is associated with small punctate structures in cultured
nonneuronal cells and in neuronal growth cones (21). Thus,
it was postulated that KIF4 is a motor for transport toward the cell
membrane, although its specific cargo in normal cells has not yet been
elucidated. Therefore, the finding of a Gag-KIF4 association suggests
that KIF4 might play a role in delivering retroviral Gag polyproteins to the plasma membrane.
Identification of proteins that interact with Pr60Gag
by use of the yeast two-hybrid system.
A cDNA library prepared
from the C57BL/Ka mouse V13 T-lymphoma cell line and cloned into the
GAL4 activation domain expression vector pACT was purchased from
Clontech (Palo Alto, Calif.). The gag genes of BM5def and
BM5eco were amplified by Vent DNA polymerase (New England Biolabs,
Beverly, Mass.) with synthetic oligonucleotide primers containing
EcoRI sites. The primers for amplification of BM5def clone
27 (2) were 5'-GATCGAATTCATGGGACAGACCATAACCAC-3' (sense) and 5'-GACTGAATTCCTAGTCACCTAAGGTTAGGA-3'
(antisense). The primers for amplification of BM5eco clone
12 (2) were 5'-GATCGAATTCATGGGACAGACCGTAACCAC-3' (sense) and 5'-GATCGAATTCCTAGTCATCTAAGGTCAGGA-3'
(antisense). The amplified products were digested with
EcoRI and cloned into the pGBT9 DNA-binding domain vector
(Clontech) to generate plasmids encoding BM5def Gag
(pGBT9-Pr60def-Gag) or BM5eco Gag
(pGBT9-Pr65eco-Gag) fusions.
To screen the V13 T-lymphoma cDNA library, Saccharomyces
cerevesiae HF7C cells carrying a GAL1-lacZ fusion gene
were first cotransformed by the lithium acetate method with
pGBT9-Pr60def-Gag and the lymphoma cDNA library. The
transformed cells were plated out on SD synthetic medium without His,
Trp, or Leu to select for cells with histidine, tryptophan, and leucine
prototropy. 
-Galactosidase activity was assayed on nitrocellulose
filter replicas of yeast transformants. Individual positive colonies were isolated, replated, and retested sequentially for
-galactosidase activity. Plasmid DNA was isolated from the blue
colonies and used to transform Escherichia coli DH5 or DH10B
(Life Technologies, Grand Island, N.Y.). Of 150,000 colonies screened,
31 true-positive clones were obtained and used to transform bacteria.
Minipreps of the 31 bacterial clones were digested with
XhoI. Three 1.4-kb inserts out of 20 different-sized
XhoI inserts were detected. One of them, which we designated
Y26, was nick translated and used as a probe in Southern blots. The
results showed that Y26 hybridized with itself and the other two
inserts of apparently identical size (data not shown).
To determine whether the protein encoded by the Y26 clone would also
react with ecotropic virus Pr65
Gag, the clone was
rescreened against yeast cells transformed with
a construct encoding
BM5eco Gag (
2) as a fusion protein with
the GAL4 DNA-binding
domain (pGBT9-Pr65
eco-Gag). Y26 reacted with both
Pr60
Gag and Pr65
Gag (data not shown). We then
selected Y26 as a candidate protein
for further study.
The Y26 insert of 1,622 bp was sequenced in its entirety and gave a
highly significant match with the GenBank sequence for
mouse KIF4
extending from bp 2577 to the end of the published
sequence
(
21) at 3976 (data not shown; KIF4 GenBank accession
no.
D12646). In the single large open reading frame (ORF) common
to both
sequences, only 2 bp distinguished the Y26 sequence from
the published
sequence for KIF4: an A for a G at 3254 and a G
for an A at 3544 of the
KIF4 sequence. Although the Y26 cDNA expressing
the Gag-binding protein
did not contain the full-length
KIF4 transcript,
the
sequence of the captured insert and the published sequence
of the
carboxy terminus of the full protein were essentially identical.
It is
unlikely that the differences seen indicate that the captured
cDNA
derives from kinesins other than KIF4, because other members
of this
protein family show the most divergence from one another
in the carboxy
terminus (
9,
12-14). We conclude that Y26 encompasses
the
carboxy terminus of KIF4.
Y26 binds to Gag polyproteins in vitro.
A GST-Y26 fusion was
constructed by subcloning the 1.5-kbp XhoI insert of Y26
into the pGEX-4T-2 vector (Pharmacia Biotechnology, Piscataway, N.J.).
The resulting construct, pGEX-4T-2-Y26, was grown in E. coli
BL21. GST-Y26 fusion protein was induced for 6 h from
BL21/pGEX-4T-2-Y26 with IPTG
(isopropyl--D-thiogalactopyranaside) and purified by
affinity binding to glutathione-Sepharose 4B according to the
procedures recommended by the manufacturer (Pharmacia Biotechnology). The GST-Y26 concentration on the beads was 2 µg/ml. The pGEX-4T-2 vector and pGEX-spectrin (kindly provided by David Bowtell, Peter MacCallum Cancer Institute, Melbourne Australia) were also grown in
E. coli BL21 to generate glutathione
S-transferase (GST) protein and GST-spectrin fusion protein
for negative controls. The concentrations of those proteins on beads
were 4 µg/ml (Fig. 1A).
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FIG. 1.
Interaction between GST-Y26 protein and ecotropic virus
Gag. (A) Protein conjugated to glutathione beads. Ten microliters of
GST-, GST-spectrin-, and GST-Y26-conjugated beads was washed,
resuspended in SDS-sample buffer, heated 10 min, and loaded on a
Tris-glycine 12% polyacrylamide gel. Coomassie blue was used for gel
staining. (B) Analysis of GST-Y26 protein-Gag association in vitro.
Twenty microliters (each) of GST-Y26-, GST-, and GST-spectrin-bound
beads was incubated with 200 µl of cell lysates for 2 h at 4 C. Precipitates were washed and loaded on a Tris-glycine 12%
polyacrylamide gel. The blot was developed with the R187 MAb. Lanes: 1, Y26 beads plus BHK-VV-eco; 2, Y26 beads plus BHK-VV; 3, 10 µl of
BHK-VV-eco lysate only; 4, GST beads plus BHK-VV-eco; 5, GST-spectrin
beads plus BHK-VV-eco.
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|
Recombinant vaccinia virus strains expressing the BM5 defective virus
gag gene or the BM5 ecotropic virus
gag gene were
constructed.
For the defective
gag gene, a 1.75-kb
gag ORF plus 120 bp of 5'
noncoding sequences isolated from
plasmid pBM5DEF27 (
2) was
ligated with a pTK-gpt-selP vector
derivative (
6,
7,
18).
For the ecotropic
gag
gene, a 1.8-kb
BanII fragment containing
the
gag
ORF from the plasmid pBM5ECO 12-1 (
2) was ligated with
a
BanII-cleaved pTZ-L2 vector derivative (
18). CV-1
cells previously
infected with wild-type vaccinia virus were
transfected with plasmid
DNAs to generate recombinant vaccinia viruses.
BHK21 cells were cultured in Dulbecco's modified Eagle's medium
(Quality Biological, Inc., Gaithersburg, Md.) containing 5%
fetal calf
serum. Cells were infected with wild-type vaccinia
virus or vaccinia
virus recombinants expressing either BM5def
or BM5eco Gag at a virus
concentration of 0.05 PFU/cell for 24
to 48 h. Cells were then
lysed in Nonidet P-40 lysis buffer (150
mM NaCl, 50 mM Tris-HCl (pH
7.0), 0.5% Nonidet P-40, 5 mM EDTA,
0.5% Tween 20, 10 µg of
aprotinin and leupeptin per ml, 1 mM phenylmethylsulfonyl
fluoride) at
a concentration of 2 × 10
7 cells/ml. Cell lysates
were precleared by overnight incubation
at 4°C with rat or rabbit
immunoglobulin G (IgG)-coupled Sepharose
beads. Total protein
concentrations were measured by the Bradford
method (Bio-Rad
Laboratories, Hercules, Calif.).
GST- or GST-Y26 fusion protein-conjugated beads were incubated with
precleared cell lysates at a ratio of 1:10 to 1:20 (vol/vol)
for 2 h at 4°C. The beads were then washed twice in buffers containing
600, 300, and 150 mM NaCl and 1 mM phenylmethylsulfonyl fluoride
individually, resuspended in 40 µl of sodium dodecyl sulfate (SDS)
reducing sample buffer, boiled for 5 min, and loaded on Tris-glycine
12% polyacrylamide minigels (Novex Experimental Technology, San
Diego,
Calif.). Separated proteins were transferred onto polyvinylidene
difluoride (PVDF) membranes. For the Western blot procedure, the
manufacturer's protocol was followed (version L.4; Tropix, Bedford,
Mass.).
The lysate of BHK21 cells infected with a vaccinia virus recombinant
expressing the BM5 ecotropic
gag gene (BHK-VV-eco) was
used
as a source of Gag polyproteins and as a positive control
(Fig.
1B,
lane 3). Pr65
Gag was detected following incubation of
GST-Y26 beads with lysates
of the BHK-VV-eco lysate (Fig.
1B, lane 1).
No Gag polyproteins
were detected following incubation of GST-Y26 beads
with lysates
of cells infected with wild-type vaccinia virus (BHK-VV)
(lane
2) or incubation of the BHK-VV-eco lysate with beads conjugated
with GST protein alone or the GST-spectrin fusion protein (lanes
4 and
5).
KIF4 binds to Gag polyproteins in vivo.
Polyclonal rabbit
anti-KIF4 antibody (21) and anti-Gag p12 monoclonal antibody
(MAb), 548, obtained from Bruce Chesebro (National Institute of Allergy
and Infectious Diseases) (5), were covalently linked to
Sepharose beads by established techniques (25) at a
concentration of 1 mg/ml. Nonimmune rabbit IgG or a rat MAb to a B-cell
surface antigen, BRB44, was also conjugated to Sepharose beads at the
same concentration and used as a negative control. Immunoprecipitations
with beads were performed with the lysates of BHK-VV-eco or BHK-VV as
described above. The amount of lysate protein used in each reaction was
about 200 µg. The blots were probed with anti-Gag p30 MAb (R187)
(Fig. 2A) or polyclonal antibody to KIF4
(Fig. 2B).
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FIG. 2.
KIF4-Gag association in BHK cells. Lysates were prepared
from BHK21 cells either uninfected (BHK) or infected with wild-type
vaccinia virus (BHK-VV) or vaccinia virus expressing ecotropic MuLV Gag
(BHK-VV-eco). Two hundred microliters of lysates containing
approximately 200 µg of cytoplasm protein was precipitated with 10 µl of Sepharose beads conjugated with polyclonal rabbit IgG anti-KIF4
(KIF4), with normal rabbit IgG (rab.IgG), with rat MAb anti-BRB44
(BRB44), or with rat anti-Gag p12 MAb (p12). Precipitated proteins were
separated on Tris-glycine 10% polyacrylamide gels and transferred to
PVDF membranes before being blotted with the indicated antibodies. IP,
immunoprecipitation.
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|
The 65-kDa Gag protein was detected in anti-KIF4 precipitates with the
lysate of BHK-VV-eco (Fig.
2A, lane 4), but not in
negative control
precipitates (lanes 2 and 3). Conversely, the
KIF4 protein with a size
of approximately 140 kDa was detected
in the anti-p12 precipitates
after incubation with the same lysate
(Fig.
2B, lane 4), but not in
control samples (lanes 2 and 3).
Normal expression of
Pr65
Gag and KIF4 was seen in lanes 1 of Fig.
2A and B,
respectively.
These results demonstrate that native KIF4 and Gag of BM5
ecotropic
virus associate in vivo when Gag is overexpressed from a
vaccinia
virus vector.
We next asked whether the KIF4-Gag association can be detected in
normal retrovirus-infected cells. To answer this question,
SC-1
(
10) and
Mus dunni (
15) cells were
infected with pools
of viruses as described previously (
20).
The SC-1 cells were
infected with ecotropic Moloney virus
(
22) or amphotropic 4070A
virus (
4), while
M. dunni cells were infected with AKR 13 mink
cytopathic
focus-forming (MCF) virus (
3) or NZB IU6 xenotropic
virus
(
3). Infected cells were harvested for biochemical analysis
on day 5 after infection. Cells were lysed and immunoprecipitated
as
described above. The Gag and KIF4 proteins were detected by
anti-KIF4
antibody or MAb R187 under the same conditions as those
described above
(Fig.
3). Pr65
Gag polyproteins were detected in lysates of
Moloney virus and 4070A
virus infected SC-1 cells precipitated with
anti-KIF4 beads (Fig.
3 [left], lanes 2 and 3) and were also in AKR 13-infected
M. dunni cells (lane
5), but not in NZB IU6-infected
M. dunni cells (lane
6). In
precipitations with R187 beads, the 140-kDa band of KIF4
was detected
in ecotropic, amphotropic, and MCF virus-infected
SC-1 or
M. dunni cells (Fig.
3 [right], lanes 2, 3, and 5), but
not in
xenotropic virus-infected cells (lane 6). The in vivo
coimmunoprecipitation
results indicate that the KIF4-Gag association
can be detected
in most retrovirus-infected cells tested in this
experiment.
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FIG. 3.
KIF4 associates with Gag in multiple MuLV-infected
cells. Lysates were prepared from uninfected SC-1 cells (lane 1) and
SC-1 cells infected with ecotropic (lane 2) or amphotropic (lane 3)
viruses and from uninfected M. dunni cells (lane 4) and
M. dunni cells infected with MCF (lane 5) or xenotropic
(lane 6) MuLV. Equivalent amounts of proteins and beads were used as
described in the legend to Fig. 2. Precipitated proteins were separated
on a Tris-glycine 12% polyacrylamide gel and transferred to PVDF
membranes before being blotted with the indicated antibodies. IP,
immunoprecipitation.
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|
KIF4 expression in mouse tissues.
If the observed KIF4-Gag
association in retrovirus-infected cells is relevant to normal
retrovirus biology, then KIF4 should be expressed ubiquitously, because
retrovirus expression can be detected in virtually all tissues of mice
that express endogenous ecotropic virus at high levels (19).
We therefore examined the expression of KIF4 in multiple mouse tissues
by using both reverse transcription-PCR and immunoblot analyses.
Because KIF4 is known to be expressed at high levels in lymphocytes, we
studied tissues from a Rag2 knockout mouse (Taconic, Germantown, N.Y.)
incapable of producing mature lymphocytes to ensure that signals for
KIF4 in lysates of different tissues did not represent infiltrating T
and B cells.
Total RNA was isolated from mouse tissues with the use of RNAzol B
(Tel-Test, Inc., Friendswood, Tex.) by the protocol of
Svetic et al.
(
23). cDNA synthesis, PCR, Southern blotting,
and
chemiluminescent detection were performed as described by
Gazzinelli et
al. (
8). For the PCRs, primers were synthesized
corresponding to the KIF4 sequence for bases 3766 to 3788 (5'-TCTTCCAGTCTCCAGACTCTTCC-3')
and the complement to bases
4396 to 4418 (5'-CTCTTCTTGGATAGGAGAGCAGC-3').
PCR conditions
were 95, then 60, and then 72°C for 60 s each for
30 cycles,
except (i) the first cycle was 180 s at 95°C, and (ii)
the last
cycle was 300 s at 72°C. The magnesium chloride concentration
was 1.25 mM. The number of cycles chosen for blot analysis was
well
short of saturation. The probe used for KIF4 detection by
hybridization
and chemiluminescence was 5'-GCCGAGCAGGACAATGAG-3'
(KIF4
bases 3894 to 3911) (
21).
Multiple tissue samples (40 to 50 mg) from a Rag2 gene-knockout mouse
and a spleen from a wild-type C57BL/6 mouse were homogenized
and lysed
in 1 ml of Nonidet P-40 lysis buffer. Postnuclear lysates
were
separated on Tris-glycine 12% polyacrylamide minigels (Novex
Experimental Technology). The blot was probed by anti-KIF4 antibody
(
21).
All tissues tested were positive both by RT-PCR (Fig.
4A) and by immunoblotting (Fig.
4B).
Thus, KIF4 is widely expressed
and would be available to participate in
virus particle formation
in a variety of tissues. This result is
apparently different from
that of a previous study, which had shown
that KIF4 transcripts
were not detectable by Northern blotting in adult
mouse tissues
other than spleen (
21). The different
techniques used for the
analysis of KIF4 expression may be responsible
for this difference.
The results reported in our study reinforce the
previous result
that KIF4 is ubiquitously expressed in almost all mouse
tissues,
although its expression level decreases during development.
This
conclusion is further supported by KIF4 expression in human
(HeLa),
monkey (Cos-7), and hamster (BHK21) cells, although expression
levels were variable (
17,
24).
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FIG. 4.
KIF4 expression in mouse tissues. (A) RT-PCR of total
RNA from the indicated tissues of a Rag2 knockout (k/o) mouse and
spleen tissue from a normal C57BL/6 (B6) mouse. (B) KIF4 protein
expression in the tissues described above. Lysates from each tissue
were loaded on a Tris-glycine 12% polyacrylamide gel, transferred to
a PVDF membrane, and then developed with anti-KIF4 antibody. ND, not
done.
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|
Our studies have shown that only a small amount of KIF4 can be
coimmunoprecipitated by anti-Gag antibodies and that only low
levels of
Gag are seen in the anti-KIF4 precipitates. This may
suggest that only
a small part of cellular KIF4 is recruited for
the transport of Gag
and/or that KIF4 rapidly dissociates from
Gag when it reaches the
plasma membrane (
16). To clarify these
issues, biochemical
characterization of the interaction of purified
KIF4 and Gag protein,
as well as its regulation, will be important.
Studies are in progress
to determine if KIF4 binds to the Gag
polyproteins of other members of
the retrovirus family. The subcellular
localization and the molecular
basis for this protein-protein
interaction are also under
investigation.
| |
ACKNOWLEDGMENTS |
The first and second authors contributed equally to this research.
We thank L. M. Lantz for expert technical advice and help; D. Lee,
T. McCarty, and D. Segal for technical assistance; and B. R. Marshall for editorial contributions.
This work was supported in part by a CRADA between IMMUNO-USA
(Rochester, Minn.) and the Laboratory of Immunopathology, National Institute of Allergy and Infectious Diseases, National Institutes of
Health, to H. C. Morse III and by a COE research grant from the
Ministry of Education, Science, and Culture of Japan to N. Hirokawa.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: LIP, NIAID, NIH,
7 Center Dr., Bethesda, MD 20892-0760. Phone: (301) 496-6379. Fax: (301) 402-0077. E-mail: hmorse{at}atlas.niaid.nih.gov.
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REFERENCES |
| 1.
|
Aziz, D. C.,
Z. Hanna, and P. Jolicoeur.
1989.
Severe immunodeficiency disease induced by a defective murine leukemia virus.
Nature
338:505-508[Medline].
|
| 2.
|
Chattopadhyay, S. K.,
D. N. Sengupta,
T. N. Fredrickson,
H. C. Morse III, and J. W. Hartley.
1991.
Characteristics and contributions of defective, ecotropic, and mink cell focus-inducing viruses involved in a retrovirus-induced immunodeficiency syndrome of mice.
J. Virol.
65:4232-4241[Abstract/Free Full Text].
|
| 3.
|
Chattopadhyay, S. K.,
M. R. Lander,
S. Gupta,
E. Rands, and D. R. Lowy.
1981.
Origin of mink cytopathic focus-forming (MCF) viruses: comparison of ecotropic and xenotropic murine leukemia virus genomes.
Virology
113:465-483[Medline].
|
| 4.
|
Chattopadhyay, S. K.,
A. I. Oliff,
D. L. Linemeyer,
M. R. Lander, and D. R. Lowy.
1981.
Genomes of murine leukemia viruses isolated from wild mice.
J. Virol.
39:777-791[Abstract/Free Full Text].
|
| 5.
|
Chesebro, B.,
W. Britt,
L. Evans,
K. Wehrly,
J. Nishio, and M. Cloyd.
1983.
Characterization of monoclonal antibodies reactive with murine leukemia viruses: use in analysis of strains of Friend MCF and Friend ecotropic murine leukemia virus.
Virology
127:134-148[Medline].
|
| 6.
|
Davison, A. J., and B. Moss.
1990.
New vaccinia virus recombination plasmids incorporating a synthetic late promoter for high level expression of foreign proteins.
Nucleic Acids Res.
18:4285-4286[Free Full Text].
|
| 7.
|
Falkner, F. G., and B. Moss.
1988.
Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors.
J. Virol.
62:1849-1854[Abstract/Free Full Text].
|
| 8.
|
Gazzinelli, R. T.,
I. Eltoum,
T. A. Wynn, and A. Sher.
1993.
Acute cerebral toxoplasmosis is induced by in vivo neutralization of TNF and correlates with the down-regulated expression of inducible nitric oxide synthase and other markers of macrophage activation.
J. Immunol.
151:3672-3681[Abstract].
|
| 9.
|
Goldstein, L. S. B.
1993.
With apologies to Scheherazade: tails of 1001 kinesin motors.
Annu. Rev. Genet.
27:319-351[Medline].
|
| 10.
|
Hartley, J. W., and W. P. Rowe.
1975.
Clonal cell lines from a feral mouse embryo which lack host-range restrictions for murine leukemia viruses.
Virology
65:128-134[Medline].
|
| 11.
|
Henderson, L. E.,
H. C. Crutzsch, and S. Oroszlan.
1983.
Myristyl amino-terminal acylation of murine retrovirus proteins: an unusual post-translational protein modification.
Proc. Natl. Acad. Sci. USA
80:339-343[Abstract/Free Full Text].
|
| 12.
|
Hirokawa, N.
1993.
Axonal transport and the cytoskeleton.
Curr. Opin. Neurobiol.
3:724-731[Medline].
|
| 13.
|
Hirokawa, N.
1996.
Organelle transport along microtubules the role of KIFS.
Trends Cell. Biol.
6:135-141.
|
| 14.
|
Hirokawa, N.
1998.
Kinesin and dynein superfamily proteins and the mechanism of organelle transport.
Science
279:519-526[Abstract/Free Full Text].
|
| 15.
|
Lander, M. R., and S. K. Chattopadhyay.
1984.
A Mus dunni cell line that lacks sequences closely related to endogenous murine leukemia viruses and can be infected by ecotropic, amphotropic, xenotropic, and mink cell focus-forming viruses.
J. Virol.
52:695-698[Abstract/Free Full Text].
|
| 16.
|
Okada, Y.,
R. Sato-Yoshitake, and N. Hirokawa.
1995.
The activation of protein kinase A pathway selectively inhibits anterograde axonal transport of vesicles but not mitochondria transport or retrograde transport in vivo.
J. Neurosci.
15:3053-3064[Abstract].
|
| 17.
| Okada, Y., and N. Hirokawa. Unpublished data.
|
| 18.
|
Pfleiderer, M.,
F. G. Falkner, and F. Dorner.
1995.
Requirements for optimal expression of secreted and nonsecreted recombinant proteins in vaccinia virus systems.
Protein Exp. Purif.
6:559-569[Medline].
|
| 19.
|
Rowe, W. P., and T. Pincus.
1972.
Quantitative studies of naturally occurring murine leukemia virus infection of AKR mice.
J. Exp. Med.
135:429-436[Abstract].
|
| 20.
|
Rowe, W. P.,
W. E. Pugh, and J. W. Hartley.
1970.
Plaque assay techniques for murine leukemia viruses.
Virology
42:1136-1139[Medline].
|
| 21.
|
Sekine, Y.,
Y. Okada,
Y. Noda,
S. Kondo,
H. Aizawa,
R. Takemura, and N. Hirokawa.
1994.
A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally.
J. Cell. Biol.
127:187-201[Abstract/Free Full Text].
|
| 22.
|
Shoemaker, C.,
S. Goff,
E. Gilboa,
M. Paskind,
S. W. Mitra, and D. Baltimore.
1980.
Structure of a cloned circular Moloney murine leukemia virus DNA molecule containing an inverted segment: implications for retrovirus integration.
Proc. Natl. Acad. Sci. USA
77:3932-3936[Abstract/Free Full Text].
|
| 23.
|
Svetic, A.,
F. D. Finkelman,
Y. C. Jian,
C. W. Dieffenbach,
D. E. Scott,
K. F. McCarthy,
A. D. Steinberg, and W. C. Gause.
1991.
Cytokine gene expression after in vivo primary immunization with goat antibody to mouse IgD antibody.
J. Immunol.
147:2391-2397[Abstract].
|
| 24.
| Tang, Y., T. A. Torrey, and H. C. Morse
III. Unpublished data.
|
| 25.
|
Vretblad, P.,
E. Hulten, and S. Bartlett.
1979.
Minimizing background adsorption to immunosorbents based on Sepharose 6MB (macrobeads), p. 797-800.
In
H. Peeters (ed.), Protides of the biological fluids, vol. 27. Pergamon Press Reprint. Franklin Book Co., Inc., Elkins Park, Pa.
|
| 26.
|
Wills, J. W., and R. C. Craven.
1991.
Form, function, and use of retroviral Gag proteins.
AIDS
5:639-654[Medline].
|
J Virol, August 1998, p. 6898-6901, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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