• CRM1
• Exportin-1
• NXF1
• Rev
• Rex
• human immunodeficiency virus
• nuclear envelope
• nuclear export
• nuclear membrane budding
• nuclear pore complex
• subcellular trafficking
• virus

• Humans
• Active Transport, Cell Nucleus
• Retroviridae/genetics
• Retroviridae/physiology
• Retroviridae/metabolism
• Genome, Viral
• Cell Nucleus/virology
• Cell Nucleus/metabolism
• Host Microbial Interactions
• Host-Pathogen Interactions
• Virus Replication
• Animals

• P01 CA022443 NCI NIH HHS
• R01 AI110221 NIAID NIH HHS
• R21 AI143800 NIAID NIH HHS
• U54 AI170660 NIAID NIH HHS
• R01AI110221 HHS | National Institutes of Health (NIH)
• P01CA022443 HHS | National Institutes of Health (NIH)
• U54AI170660 Greater Milwaukee Foundation (GMF)

• CRM1
• NPC
• NXF1
• Nup98
• Rae1
• host mRNA
• mRNA export
• viral mRNA
• virus

• Humans
• Active Transport, Cell Nucleus
• Virus Diseases
• Antiviral Agents
• RNA Transport
• Eukaryota
• RNA, Messenger/genetics

• 22PJ1414600 Science and Technology Commission of Shanghai Municipality
• 21PJ1414500 Science and Technology Commission of Shanghai Municipality
• HS2021SHZX001 Science and Technology Commission of Shanghai Municipality

• RNA export
• TREX-TAP pathway
• cytoplasmic accumulation region
• m6A modification
• nuclear retention
• nuclear retention element

• 31670823 National Natural Science Foundation of China
• 32171290 National Natural Science Foundation of China

This review is an accompaniment to a Special Issue on “Retroviral RNA Processing”. It discusses post-transcriptional regulation of retroviruses, ranging from the ancient foamy viruses to more modern viruses, such as HIV-1, HTLV-1, Rous sarcoma virus, murine leukemia virus, mouse mammary tumor virus, and Mason-Pfizer monkey virus. This review is not comprehensive. However, it tries to address some of the major questions in the field with examples of how different retroviruses express their genes. It is amazing that a single primary RNA transcript can have so many possible fates: genomic RNA, unspliced mRNA, and up to 50 different alternatively spliced mRNAs. This review will discuss the sorting of RNAs for packaging or translation, RNA nuclear export mechanisms, splicing, translation, RNA modifications, and avoidance of nonsense-mediated RNA decay.

• NMD
• RNA modification
• RNA processing
• export
• foamy viruses
• retroviruses
• splicing
• translation

• Active Transport, Cell Nucleus
• Animals
• Leukemia Virus, Murine/genetics
• Mice
• RNA Processing, Post-Transcriptional
• RNA, Messenger/metabolism
• RNA, Viral/genetics
• RNA, Viral/metabolism
• Retroviridae/genetics
• Retroviridae/metabolism

• nucleus
• viral RNA nuclear export
• viral ribonucleoprotein complexes
• virus

• Active Transport, Cell Nucleus/physiology
• Cell Nucleus/virology
• Cytoplasm/metabolism
• DNA Viruses/physiology
• Humans
• Karyopherins/metabolism
• Nucleocytoplasmic Transport Proteins/metabolism
• RNA Transport
• RNA, Messenger
• RNA, Viral/genetics
• RNA, Viral/physiology
• Retroviridae/physiology
• Viral Proteins/metabolism

• CTEs
• RNA Export
• RNA splicing
• alternative splicing
• intron retention
• nucelocytoplasmic export

• Active Transport, Cell Nucleus/physiology
• Cell Nucleus/metabolism
• Gene Expression Regulation, Viral
• Gene Products, rev/metabolism
• Genes, env/physiology
• HIV-1
• Mason-Pfizer monkey virus
• Molecular Imaging/methods
• RNA Processing, Post-Transcriptional/physiology
• RNA, Messenger/metabolism
• RNA, Viral
• Regulatory Sequences, Nucleic Acid/genetics
• Regulatory Sequences, Nucleic Acid/physiology
• Regulatory Sequences, Ribonucleic Acid/genetics
• Regulatory Sequences, Ribonucleic Acid/physiology
• Single-Cell Analysis/methods

• P01 CA022443 NCI NIH HHS
• R01 AI110221 NIAID NIH HHS
• T32 CA009135 NCI NIH HHS

The nonsense-mediated mRNA decay (NMD) pathway degrades mRNAs containing long 3'UTRs to perform dual roles in mRNA quality control and gene expression regulation. However, expansion of vertebrate 3'UTR functions has required a physical expansion of 3'UTR lengths, complicating the process of detecting nonsense mutations. We show that the polypyrimidine tract binding protein 1 (PTBP1) shields specific retroviral and cellular transcripts from NMD. When bound near a stop codon, PTBP1 blocks the NMD protein UPF1 from binding 3'UTRs. PTBP1 can thus mark specific stop codons as genuine, preserving both the ability of NMD to accurately detect aberrant mRNAs and the capacity of long 3'UTRs to regulate gene expression. Illustrating the wide scope of this mechanism, we use RNA-seq and transcriptome-wide analysis of PTBP1 binding sites to show that many human mRNAs are protected by PTBP1 and that PTBP1 enrichment near stop codons correlates with 3'UTR length and resistance to NMD.

DOI:http://dx.doi.org/10.7554/eLife.11155.001

Genes are used as templates to create molecules of messenger RNA (mRNA) that contain all the information needed to make a protein. This information begins with a 'start site' and ends with a 'stop site.' The regions of the mRNA outside of the start and stop sites are called untranslated regions.

Not all mRNAs are correctly made, and cells combat this problem by detecting and destroying faulty mRNAs before they are translated into protein. One way cells do this is by recognizing and destroying mRNAs that include long untranslated regions, which can indicate that the mRNA might have a stop site too early in its sequence. A key problem with this mechanism, however, is that long untranslated regions also serve important roles in the cell: for example, by determining where and when mRNA molecules are read to make protein. How then do mRNAs with long but important untranslated regions escape detection and degradation?

Ge et al. have now investigated this question using an approach that allows a 'handle' to be attached to particular RNA molecules. This allows the RNA and any proteins bound to it to be purified away from all other RNAs and proteins in the cell, and the proteins can then be identified by a technique called mass spectrometry.

Ge at al. found that mRNAs can recruit a protein called PTBP1 to part of the RNA sequence near the stop site. This prevents an RNA decay protein recognizing and triggering the degradation of the mRNA, even if the mRNA has a long untranslated region.

Thus, PTBP1 plays a crucial role in protecting human RNAs with long untranslated regions from destruction by the nonsense-mediated decay pathway. Some viral RNAs are also able to evade decay, and so Ge et al. hypothesize that the virus stole this method for maintaining its RNAs from host cells. A future goal is to understand whether this system works the same way in all cell types or protects different RNAs in different cells.

DOI:http://dx.doi.org/10.7554/eLife.11155.002

• 3'UTR
• PTBP1
• Rous sarcoma virus
• biochemistry
• chicken
• human
• infectious disease
• microbiology
• nonsense-mediated mRNA decay
• retrovirus

The UL49 ORF of human cytomegalovirus (HCMV) is essential for viral replication; conserved among all herpes viruses; however, the function is unclear. Once the UL49 ORF was precisely deleted from the start to stop codon, the mutant did not yield infectious progeny. In this study, we find out many alternatively processed ESTs in UL49 locus in HCMV-infected cells, in which there are two novel transcription termination sites in UL49 locus. Most of these ESTs are rare transcripts that contain directed repeat sequences in the intron splicing regions. There is a typical GU-AG intron splicing site in UL49Y transcripts. The 1847 bp UL49Y cDNA spans an ORF from 335 to 1618 and encodes a putative protein of 427 amino acids with a predicted molecular mass of 47.1 kDa. All the new EST sequences and UL49Y cDNA sequence have been deposited in the GenBank database (GenBank Accession nos. GW314860-GW314900 and GU376796). This study provides us with very important clues for revealing the importance of the UL49 locus alternative splicing.

All retroviruses synthesize essential proteins via alternatively spliced mRNAs. Retrovirus genera, though, exploit different mechanisms to coordinate the synthesis of proteins from alternatively spliced mRNAs. The best studied of these retroviral, post-transcriptional effectors are the trans-acting Rev protein of lentiviruses and the cis-acting constitutive transport element (CTE) of the betaretrovirus Mason-Pfizer monkey virus (MPMV). How members of the gammaretrovirus genus translate protein from unspliced RNA has not been elucidated.

The mechanism by which two gammaretroviruses, XMRV and MLV, synthesize the Gag polyprotein (Pr65^Gag) from full-length, unspliced mRNA was investigated here. The yield of Pr65^Gag from a gag–only expression plasmid was found to be at least 30-fold less than that from an otherwise isogenic gag-pol expression plasmid. A frameshift mutation disrupting the pol open reading frame within the gag-pol expression plasmid did not decrease Pr65^Gag production and 398 silent nucleotide changes engineered into gag rendered Pr65^Gag synthesis pol-independent. These results are consistent with pol-encoded RNA acting in cis to promote Pr65^Gag translation. Two independently-acting pol fragments were identified by screening 17 pol deletion mutations. To determine the mechanism by which pol promoted Pr65^Gag synthesis, gag RNA in total and cytoplasmic fractions was quantitated by northern blot and by RT-PCR. The pol sequences caused, maximally, three-fold increase in total or cytoplasmic gag mRNA. Instead, pol sequences increased gag mRNA association with polyribosomes ~100-fold, a magnitude sufficient to explain the increase in Pr65^Gag translation efficiency. The MPMV CTE, an NXF1-binding element, substituted for pol in promoting Pr65^Gag synthesis. A pol RNA stem-loop resembling the CTE promoted Pr65^Gag synthesis. Over-expression of NXF1 and NXT, host factors that bind to the MPMV CTE, synergized with pol to promote gammaretroviral gag RNA loading onto polysomes and to increase Pr65^Gag synthesis. Conversely, Gag polyprotein synthesis was decreased by NXF1 knockdown. Finally, overexpression of SRp20, a shuttling protein that binds to NXF1 and promotes NXF1 binding to RNA, also increased gag RNA loading onto polysomes and increased Pr65^Gag synthesis.

These experiments demonstrate that gammaretroviral pol sequences act in cis to recruit NXF1 and SRp20 to promote polysome loading of gag RNA and, thereby license the synthesis of Pr65^Gag from unspliced mRNA.

Post-transcriptional regulatory mechanisms of several complex and simple retroviruses and retroelements have been elucidated, with the exception of the gammaretrovirus family. We found that, similar to the other retroviruses, gag gene expression of MuLV and XMRV depends on post-transcriptional regulation mediated via an RNA sequence overlapping the pro-pol open reading frame, termed the Post-Transcriptional Element (PTE). PTE function can be replaced by heterologous RNA export elements, e.g. CTE of simian type D retroviruses. Alternatively, Gag particle production is achieved using an RNA/codon optimized gag gene. PTE function is transferable and can replace HIV Rev-RRE-regulated expression of HIV gag. Analysis of PTE by SHAPE revealed a highly structured RNA comprising seven stem-loop structures, with the 5′ and 3′ stem-loops forming an essential bipartite signal. MuLV and XMRV PTE share 98% identity and have highly similar RNA structures, with changes mostly located to single-stranded regions. PTE identification strongly suggests that all retroviruses and retroelements share common strategies of post-transcriptional gene regulation to produce Gag. Expression depends on complex RNA structures embedded within retroviral mRNA, in coding regions or the 3′ untranslated region. These specific structures serve as recognition signals for either cellular or viral proteins.

AIM: To search for the presence of cis elements in hepatitis D virus (HDV) genomic and antigenomic RNA capable of promoting nuclear export.

METHODS: We made use of a well characterized chloramphenicol acetyl-transferase reporter system based on plasmid pDM138. Twenty cDNA fragments corresponding to different HDV genomic and antigenomic RNA sequences were inserted in plasmid pDM138, and used in transfection experiments in Huh7 cells. The relative amounts of HDV RNA in nuclear and cytoplasmic fractions were then determined by real-time polymerase chain reaction and Northern blotting. The secondary structure of the RNA sequences that displayed nuclear export ability was further predicted using a web interface. Finally, the sensitivity to leptomycin B was assessed in order to investigate possible cellular pathways involved in HDV RNA nuclear export.

RESULTS: Analysis of genomic RNA sequences did not allow identifying an unequivocal nuclear export element. However, two regions were found to promote the export of reporter mRNAs with efficiency higher than the negative controls albeit lower than the positive control. These regions correspond to nucleotides 266-489 and 584-920, respectively. In addition, when analyzing antigenomic RNA sequences a nuclear export element was found in positions 214-417. Export mediated by the nuclear export element of HDV antigenomic RNA is sensitive to leptomycin B suggesting a possible role of CRM1 in this transport pathway.

CONCLUSION: A cis-acting nuclear export element is present in nucleotides 214-417 of HDV antigenomic RNA.

• Hepatitis D virus
• Genomic RNA
• Antigenomic RNA
• Nuclear export
• Nuclear export element

• retrotransposon
• gag
• mrna
• nuclear export
• ty1
• saccharomyces

• Active Transport, Cell Nucleus
• Adaptor Proteins, Signal Transducing/genetics
• Cell Nucleus/metabolism
• Cytoplasm/metabolism
• Exosomes/metabolism
• Frameshift Mutation
• Fusion Proteins, gag-pol/genetics
• Fusion Proteins, gag-pol/metabolism
• Gene Deletion
• RNA Stability
• RNA Transport
• RNA, Messenger/metabolism
• Retroelements/genetics
• Retroviridae/metabolism
• Saccharomyces cerevisiae/genetics
• Saccharomyces cerevisiae/metabolism
• Saccharomyces cerevisiae Proteins/genetics

• GM095622 NIGMS NIH HHS
• R01 GM095622 NIGMS NIH HHS
• Intramural NIH HHS
• HHSN261200800001E NCI NIH HHS
• HHSN2612008000001E PHS HHS
• HHSN261200800001C NCI NIH HHS

Jaagsiekte sheep retrovirus (JSRV) is the causative agent of a lung cancer in sheep known as ovine pulmonary adenocarcinoma (OPA). The disease has been identified around the world in several breeds of sheep and goats, and JSRV infection typically has a serious impact on affected flocks. In addition, studies on OPA are an excellent model for human lung carcinogenesis. A unique feature of JSRV is that its envelope (Env) protein functions as an oncogene. The JSRV Env-induced transformation or oncogenesis has been studied in a variety of cell systems and in animal models. Moreover, JSRV studies have provided insights into retroviral genomic RNA export/expression mechanisms. JSRV encodes a trans-acting factor (Rej) within the env gene necessary for the synthesis of Gag protein from unspliced viral RNA. This review summarizes research pertaining to JSRV-induced pathogenesis, Env transformation, and other aspects of JSRV biology.

• Env
• JSRV
• Jaagsiekte sheep retrovirus
• Rej
• cancer
• ovine pulmonary adenocarcinoma
• transformation

Nonsense-mediated mRNA decay (NMD) is an mRNA quality control mechanism that selectively recognizes and targets for degradation mRNAs containing premature termination codons. Retroviral full-length RNA is presented to the host translation machinery with characteristics rarely observed among host cell mRNAs: a long 3' UTR, retained introns, and multiple open reading frames. As a result, the viral RNA is predicted to be recognized by the host NMD machinery and degraded. In the case of the Rous sarcoma virus (RSV), we identified a stability element (RSE), which resides immediately downstream of the gag termination codon and facilitates NMD evasion.

We defined key RNA features of the RSE through directed mutagenesis of the virus. These data suggest that the minimal RSE is 155 nucleotides (nts) and functions independently of the nucleotide sequence of the stop codon or the first nucleotide following the stop codon. Further data suggested that the 3'UTRs of the RSV pol and src may also function as stability elements.

We propose that these stability elements in RSV may be acting as NMD insulators to mask the preceding stop codon from the NMD machinery.

Once transcribed, the nascent full-length RNA of HIV-1 must travel to the appropriate host cell sites to be translated or to find a partner RNA for copackaging to form newly generated viruses. In this report, we sought to delineate the location where HIV-1 RNA initiates dimerization and the influence of the RNA transport pathway used by the virus on downstream events essential to viral replication. Using a cell-fusion-dependent recombination assay, we demonstrate that the two RNAs destined for copackaging into the same virion select each other mostly within the cytoplasm. Moreover, by manipulating the RNA export element in the viral genome, we show that the export pathway taken is important for the ability of RNA molecules derived from two viruses to interact and be copackaged. These results further illustrate that at the point of dimerization the two main cellular export pathways are partially distinct. Lastly, by providing Gag in trans, we have demonstrated that Gag is able to package RNA from either export pathway, irrespective of the transport pathway used by the gag mRNA. These findings provide unique insights into the process of RNA export in general, and more specifically, of HIV-1 genomic RNA trafficking.

High genetic diversity of HIV-1 presents a difficult barrier for drug treatment and vaccine development. HIV-1 efficiently reassorts mutations in its genome by packaging two copies of RNA into one virion and using portions of each RNA as the template for DNA synthesis. Thus, a key to HIV-1's genetic diversity is how it packages two RNAs into one virion. We have previously shown that HIV-1 packages RNA as a dimer, rather than two monomers. In this report, we determined that HIV-1 RNA finds its copackaged dimer partner mostly in the cytoplasm, not the nucleus, of the infected cells. We also found that the host cell machinery used to transport the HIV-1 RNA out of the nucleus affects the ability of two RNAs to copackage into one virion: HIV-1 RNAs transported using the CRM-1 pathway does not copackage efficiently with RNA transported by the NXF1 pathway. This is also the first demonstration that RNAs transported by the two major cellular pathways, CRM-1 and NXF1, are partially segregated at some point in the cytoplasm. Therefore, this work provides novel insights to both HIV replication mechanisms and cellular RNA transport pathways.

• rna export
• rous sarcoma virus
• tap
• dbp5
• retrovirus

• Animals
• Cells, Cultured
• Cytoplasm/metabolism
• Gene Expression Regulation, Viral
• Gene Products, gag/physiology
• Mutation
• Nucleocytoplasmic Transport Proteins/physiology
• RNA Splicing
• RNA, Viral/metabolism
• Repetitive Sequences, Nucleic Acid/genetics
• Rous sarcoma virus/genetics
• Rous sarcoma virus/physiology
• Sarcoma, Avian/metabolism
• Sarcoma, Avian/virology
• Transcription Factors/physiology

• R01 CA048746 NCI NIH HHS
• T32 GM007231 NIGMS NIH HHS
• T32GM07231 NIGMS NIH HHS
• R01 CA48746 NCI NIH HHS

• Animals
• Base Sequence
• Gene Expression Regulation, Viral
• HIV-1/genetics
• Humans
• Molecular Sequence Data
• RNA, Viral/chemistry
• RNA, Viral/genetics
• Retroviridae/genetics
• Terminal Repeat Sequences/genetics
• Viral Proteins/metabolism

• Alternative Splicing
• Animals
• Cell Nucleus/virology
• Cytoplasm/virology
• Humans
• RNA Processing, Post-Transcriptional
• RNA Transport
• RNA, Messenger/genetics
• RNA, Viral/genetics
• Retroviridae/genetics
• Retroviridae/growth & development
• Viral Proteins/genetics

• R01 CA078709 NCI NIH HHS
• R01 CA78709 NCI NIH HHS

• Animals
• Cell Nucleus/chemistry
• Cell Nucleus/metabolism
• Cytoplasm/chemistry
• Cytoplasm/metabolism
• Fatty Acids, Unsaturated/chemistry
• Fatty Acids, Unsaturated/pharmacology
• Humans
• Karyopherins/physiology
• Models, Biological
• Protein Transport/physiology
• Receptors, Steroid/antagonists & inhibitors
• Receptors, Steroid/metabolism
• Signal Transduction/drug effects
• Signal Transduction/physiology

• 5' Untranslated Regions
• Active Transport, Cell Nucleus
• Animals
• Capsid/metabolism
• Cell Fractionation
• Cell Nucleus/metabolism
• Cytoplasm/metabolism
• Dimerization
• Fluorescent Dyes
• In Situ Hybridization, Fluorescence
• Indoles
• Introns
• Mice
• Microscopy, Fluorescence
• Moloney murine leukemia virus/chemistry
• Moloney murine leukemia virus/genetics
• Mutation
• NIH 3T3 Cells
• Nucleic Acid Conformation
• RNA Splicing
• RNA, Ribosomal/analysis
• RNA, Ribosomal/chemistry
• RNA, Small Nucleolar/analysis
• RNA, Small Nucleolar/metabolism
• RNA, Transfer/analysis
• RNA, Transfer/chemistry
• RNA, Viral/analysis
• RNA, Viral/chemistry
• RNA, Viral/isolation & purification
• RNA, Viral/metabolism
• Transfection
• Virion/chemistry
• Virion/genetics
• Virus Replication/genetics

• 5' Untranslated Regions
• Animals
• Cell Line
• Cytomegalovirus/genetics
• Gene Expression
• Genes, Regulator
• Genetic Vectors
• Green Fluorescent Proteins/genetics
• HSP70 Heat-Shock Proteins/genetics
• HeLa Cells
• Humans
• Leukemia Virus, Murine/genetics
• Marmota
• Mason-Pfizer monkey virus/genetics
• Mice
• Muromegalovirus/genetics
• NIH 3T3 Cells
• Plasmids/genetics
• Recombinant Proteins/genetics
• Retroviridae/genetics
• Retroviridae/physiology
• Virus Replication

• Animals
• Base Sequence
• Genome
• Humans
• Mice
• Molecular Sequence Data
• Nucleic Acid Conformation
• Oocytes
• RNA/analysis
• RNA/chemistry
• RNA Transport/genetics
• Sequence Homology, Nucleic Acid
• Xenopus laevis

• Animals
• Cell Transformation, Neoplastic/genetics
• Cell Transformation, Neoplastic/metabolism
• Gene Expression Regulation, Viral/genetics
• HIV-1/genetics
• HIV-1/metabolism
• Herpesviridae Infections/genetics
• Herpesviridae Infections/metabolism
• Herpesvirus 2, Saimiriine/genetics
• Herpesvirus 2, Saimiriine/metabolism
• Humans
• Lymphoma, T-Cell/genetics
• Lymphoma, T-Cell/metabolism
• Models, Biological
• Oncogene Proteins, Viral/biosynthesis
• Oncogene Proteins, Viral/genetics
• Tumor Virus Infections/genetics
• Tumor Virus Infections/metabolism

Most mRNA-encoding genes require introns for efficient expression in high eukaryotes. However, mRNAs can efficiently accumulate in the cytoplasm without intron excision if they contain cis-acting elements such as the post-transcriptional regulatory element (PRE) of hepatitis B virus (HBV), the constitutive transport element (CTE) of Mason–Pfizer monkey virus (MPMV), or the pre-mRNA processing enhancer (PPE) of herpes simplex virus' thymidine kinase (HSV-TK) gene. We compared the activities of these viral elements, the Rev-responsive element (RRE) of the human immunodeficiency virus (HIV), and the human c-Jun gene's enhancer (CJE), an element newly identified here, to enable expression of an intronless variant of the human β-globin gene. The PRE, PPE and CJE from naturally intronless genes, but not the CTE or RRE from intron-containing genes, significantly enhanced stability, 3′ end processing and cytoplasmic accumulation. When the transcripts included the β-globin gene's first intron, the PRE, PPE and CJE still enhanced mRNA biogenesis, in some cases without intron excision. Thus, elements enabling stability, 3′ end formation and nucleocytoplasmic export, not the presence of introns or their excision per se, are necessary for mRNA biogenesis. While the CTE and RRE primarily enhance nucleocytoplasmic export, PPE-like elements from naturally intronless genes facilitate polyadenylation as well.

• Active Transport, Cell Nucleus
• Animals
• COS Cells
• Cell Line
• Cell Nucleus/metabolism
• Chlorocebus aethiops
• Cytoplasm/metabolism
• Genes, jun
• Globins/biosynthesis
• Globins/genetics
• Humans
• Introns
• Polyadenylation
• RNA 3' End Processing
• RNA Precursors/metabolism
• RNA Stability
• RNA, Messenger/biosynthesis
• RNA, Messenger/metabolism
• RNA, Viral/chemistry
• Regulatory Sequences, Ribonucleic Acid
• Transcription, Genetic

• P01 CA022443 NCI NIH HHS
• P30 CA014520 NCI NIH HHS
• CA014520 NCI NIH HHS
• CA22443 NCI NIH HHS

• Animals
• Cell Line
• Gene Expression
• Genetic Therapy/methods
• Genetic Vectors/administration & dosage
• Hematopoietic Stem Cells
• Humans
• Infectious Anemia Virus, Equine/genetics
• Peptide Elongation Factor 1/genetics
• Promoter Regions, Genetic
• Transduction, Genetic/methods
• Transgenes
• Virus Inactivation

• AI47693 NIAID NIH HHS
• HL10430 NHLBI NIH HHS

• Avian Sarcoma Viruses/genetics
• Gene Products, gag/physiology
• Protein Biosynthesis
• RNA Splicing
• RNA, Messenger/metabolism
• RNA, Viral/genetics
• Virus Assembly

• R01 CA048746 NCI NIH HHS
• T32 GM007231 NIGMS NIH HHS
• R01 CA 48746 NCI NIH HHS
• T32 GM 07231 NIGMS NIH HHS

• 5' Untranslated Regions/chemistry
• Gene Products, gag/biosynthesis
• Gene Products, gag/genetics
• Genes, Reporter
• HIV-1/genetics
• Humans
• Nucleic Acid Conformation
• Protein Biosynthesis
• RNA Splicing
• RNA, Viral/chemistry
• Retroviridae/genetics
• Terminal Repeat Sequences

• P30 CA016058 NCI NIH HHS
• P30CA16058 NCI NIH HHS
• R29A140851 PHS HHS

• AIDS Vaccines/administration & dosage
• AIDS Vaccines/immunology
• Animals
• Cytokines/biosynthesis
• Cytotoxicity Tests, Immunologic
• Enhancer Elements, Genetic/genetics
• Female
• Gene Products, rev/genetics
• HIV Antibodies/blood
• HIV Envelope Protein gp160/genetics
• HIV Envelope Protein gp160/immunology
• HIV Infections/immunology
• HIV Infections/prevention & control
• HIV-1/classification
• HIV-1/genetics
• HIV-1/immunology
• Humans
• Lymphocyte Activation
• Mason-Pfizer monkey virus/genetics
• Mice
• Mice, Inbred BALB C
• Plasmids/genetics
• T-Lymphocytes/immunology
• T-Lymphocytes, Cytotoxic
• Vaccines, DNA/administration & dosage
• Vaccines, DNA/immunology
• rev Gene Products, Human Immunodeficiency Virus

• Active Transport, Cell Nucleus
• Animals
• Cell Nucleus/metabolism
• Gene Products, rev/metabolism
• HIV-1/physiology
• Models, Biological
• RNA, Messenger/metabolism
• RNA, Viral
• rev Gene Products, Human Immunodeficiency Virus

• Cell Nucleus/metabolism
• Gene Products, env/physiology
• HIV-1/chemistry
• HIV-1/physiology
• HeLa Cells
• Humans
• Nuclear Localization Signals
• RNA, Viral/metabolism
• Virus Assembly

• Animals
• Cell Line
• Cell Line, Transformed
• Gene Expression Regulation/physiology
• Gene Transfer Techniques
• Genes, Regulator/genetics
• Genetic Vectors
• Green Fluorescent Proteins
• HIV-1/genetics
• Humans
• Infectious Anemia Virus, Equine/genetics
• Lentivirus/genetics
• Luminescent Proteins/genetics
• Luminescent Proteins/metabolism
• Mice
• Muscle, Skeletal/metabolism
• RNA Stability
• RNA, Messenger
• Transduction, Genetic
• Tumor Cells, Cultured

• AI47693 NIAID NIH HHS
• HL10430 NHLBI NIH HHS

• Animals
• Genome
• Humans
• Long Interspersed Nucleotide Elements
• RNA Interference
• RNA Transport
• Short Interspersed Nucleotide Elements