scholarly journals Retroviral targeting of proliferating endothelial cells.

2005 ◽  
Vol 52 (3) ◽  
pp. 731-735 ◽  
Author(s):  
Alexander Gornikiewicz ◽  
Anna Zommer ◽  
Raimund Jakesz ◽  
Michael Gnant ◽  
Christine Brostjan
Keyword(s):  
Endothelial Cells ◽  
Tumor Tissue ◽  
Leukemia Virus ◽  
Chimeric Protein ◽  
Retroviral Vectors ◽  
Genetically Engineered ◽  
Moloney Murine Leukemia Virus ◽  
Human Endothelial Cells ◽  
Virus Envelope ◽  
Surface Molecules

Tumor growth requires the formation of new blood vessels by endothelial cells. Thus, surface molecules -- such as angiogenin receptors -- that are selectively expressed on growing endothelium represent an attractive target for directed delivery of compounds to tumor tissue. We attempted to obtain genetically engineered retroviral vectors targeted to the endothelium by inserting the human angiogenin sequence into Moloney murine leukemia virus envelope glycoprotein. Abundant expression of the chimeric protein could be verified. However, while being selective for proliferating human endothelial cells, the recombinant retroviral particles displayed low transduction efficiencies and thus have to be further improved.

scholarly journals Sequential activation of the three protomers in the Moloney murine leukemia virus Env

2017 ◽  
Vol 114 (10) ◽  
pp. 2723-2728 ◽  
Author(s):  
Mathilda Sjöberg ◽  
Robin Löving ◽  
Birgitta Lindqvist ◽  
Henrik Garoff
Keyword(s):  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Moloney Murine Leukemia Virus ◽  
Virus Envelope ◽  
Sequential Activation ◽  
Virus Envelope Protein ◽  
Membrane Fusion Proteins ◽  
Viral Membrane Fusion ◽  
Murine Leukemia

Viral membrane fusion proteins of class I are trimers in which the protomeric unit is a complex of a surface subunit (SU) and a fusion active transmembrane subunit (TM). Here we have studied how the protomeric units of Moloney murine leukemia virus envelope protein (Env) are activated in relation to each other, sequentially or simultaneously. We followed the isomerization of the SU-TM disulfide and subsequent SU release from Env with biochemical methods and found that this early activation step occurred sequentially in the three protomers, generating two asymmetric oligomer intermediates according to the scheme (SU-TM)3→ (SU-TM)2TM → (SU-TM)TM2→ TM3. This was the case both when activation was triggered in vitro by depleting stabilizing Ca2+from solubilized Env and when viral Env was receptor triggered on rat XC cells. In the latter case, the activation reaction was too fast for direct observation of the intermediates, but they could be caught by alkylation of the isomerization active thiol.

scholarly journals Role of intron-contained sequences in formation of moloney murine leukemia virus env mRNA.

1984 ◽  
Vol 4 (11) ◽  
pp. 2289-2297 ◽  
Author(s):  
L S Hwang ◽  
J Park ◽  
E Gilboa
Keyword(s):  
Splice Site ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Splice Junction ◽  
Moloney Murine Leukemia Virus ◽  
Expression Vectors ◽  
Virus Envelope ◽  
Cellular Genes ◽  
Murine Leukemia

Formation of the Moloney murine leukemia virus envelope mRNA involves the removal of a 5,185-base pair-long intron. Deletion analysis of two Moloney murine leukemia virus-derived expression vectors revealed the existence of two short regions within the viral intron which are required for the efficient formation of the spliced RNA species. One region was present upstream from the 3' splice junction, extended at least 85 nucleotides beyond the splice site, and was not more than 165 nucleotides long. As yeast polymerase II introns, the Moloney murine leukemia virus intron contains the sequence 5'-TACTAAC-3' 15 nucleotides upstream from the 3' splice site. A second region located in the middle of the intron, within a 560-nucleotide-long sequence, was also essential for formation of the spliced RNA species. The efficient splicing of the env mRNA in the absence of expression of viral genes raises the possibility that similar mechanisms are used to remove introns of (some) cellular genes.

The in vitro interaction of Sporothrix schenckii with human endothelial cells is modulated by cytokines and involves endothelial surface molecules

2004 ◽  
Vol 36 (4) ◽  
pp. 177-188 ◽  
Author(s):  
Camila Castro Figueiredo ◽  
Osana Cunha de Lima ◽  
Laı́s de Carvalho ◽  
Leila Maria Lopes-Bezerra ◽  
Verônica Morandi
Keyword(s):  
Endothelial Cells ◽  
Sporothrix Schenckii ◽  
Human Endothelial Cells ◽  
Surface Molecules ◽  
Endothelial Surface ◽  
In Vitro Interaction

scholarly journals Moloney Murine Leukemia Virus Envelope Protein Subunits, gp70 and Pr15E, Form a Stable Disulfide-Linked Complex

1998 ◽  
Vol 72 (8) ◽  
pp. 6537-6545 ◽  
Author(s):  
Dirk-Jan E. Opstelten ◽  
Michael Wallin ◽  
Henrik Garoff
Keyword(s):  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Alkylating Agents ◽  
Moloney Murine Leukemia Virus ◽  
Virus Envelope ◽  
Disulfide Linkage ◽  
Virus Envelope Protein ◽  
Biochemical Analyses ◽  
Murine Leukemia

ABSTRACT The nature and stability of the interactions between the gp70 and Pr15E/p15E molecules of murine leukemia virus (MLV) have been disputed extensively. To resolve this controversy, we have performed quantitative biochemical analyses on gp70-Pr15E complexes formed after independent expression of the amphotropic and ecotropic Moloney MLVenv genes in BHK-21 cells. We found that all cell-associated gp70 molecules are disulfide linked to Pr15E whereas only a small amount of free gp70 is released by the cells. The complexes were resistant to treatment with reducing agents in vivo, indicating that the presence and stability of the disulfide interaction between gp70 and Pr15E are not dependent on the cellular redox state. However, disulfide-bonded Env complexes were disrupted in lysates of nonalkylated cells in a time-, temperature-, and pH-dependent fashion. Disruption seemed not to be caused by a cellular factor but is probably due to a thiol-disulfide exchange reaction occurring within the Env complex after solubilization. The possibility that alkylating agents induce the formation of the intersubunit disulfide linkage was excluded by showing that disulfide-linked gp70-Pr15E complexes exist in freshly made lysates of nonalkylated cells and that disruption of the complexes can be prevented by lowering the pH. Together, these data establish that gp70 and Pr15E form a stable disulfide-linked complex in vivo.

scholarly journals Construction and properties of replication-competent murine retroviral vectors encoding methotrexate resistance.

1989 ◽  
Vol 9 (1) ◽  
pp. 100-108 ◽  
Author(s):  
H Stuhlmann ◽  
R Jaenisch ◽  
R C Mulligan
Keyword(s):  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Retroviral Vectors ◽  
Moloney Murine Leukemia Virus ◽  
Drug Selection ◽  
Cdna Insert ◽  
Methotrexate Resistance ◽  
Foreign Genes ◽  
Murine Leukemia

A series of replication-competent Moloney murine leukemia virus vectors was constructed in which each vector contained a mutant dihydrofolate reductase (DHFR) cDNA insert in the U3 region of the viral long terminal repeat. Two of the resulting viruses, MLV (murine leukemia virus) DHFR*-5 and MLV DHFR*-7, were able to stably transfer methotrexate resistance to infected fibroblast cells upon multiple rounds of virus replication and in the absence of drug selection. Cell lines producing recombinant virus with high titers were established, which indicated that the insert did not grossly interfere with viral replication functions. These vectors should be useful for introducing and expressing foreign genes in vivo in tissues and whole animals in which virus spread is needed for efficient infection.

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