Poly(A) site selection in the HIV-1 provirus: inhibition of promoter-proximal polyadenylation by the downstream major splice donor site.

In common with all retroviruses, the human immunodeficiency virus type 1 (HIV-1) contains duplicated long terminal repeat (LTR) sequences flanking the proviral genome. These LTRs contain identical poly(A) signals, which are both transcribed into RNA. Therefore, to allow efficient viral expression, a mechanism must exist to either restrict promoter-proximal poly(A) site use or enhance the activity of the promoter-distal poly(A) site. We have examined the use of both poly(A) sites using proviral clones. Mutation of the previously defined upstream activatory sequences of the 3' LTR poly(A) site decreases the efficiency of polyadenylation when placed in competition with an efficient downstream processing signal. However, in the absence of competition, these mutations have no effect on HIV-1 polyadenylation. In addition, the 5' LTR poly(A) site is inactive, whereas a heterologous poly(A) site positioned in its place is utilized efficiently. Furthermore, transcription initiating from the 3' LTR promoter utilizes the 3' LTR poly(A) signal efficiently. Therefore, the main determinant of the differential poly(A) site use appears to be neither proximity to a promoter element in the 5' LTR nor the presence of upstream activating sequences at the 3' LTR. Instead, we show that the major splice donor site that is immediately downstream of the 5' LTR inhibits cleavage and polyadenylation at the promoter-proximal site. The fact that this poly(A) site is active in a proviral clone when the major splice donor site is mutated suggests that the selective use of poly(A) signals in HIV-1 is mediated by a direct inhibition of the HIV-1 poly(A) site by downstream splicing events or factors involved in splicing.

[1]  M. Wollerton,et al.  Upstream sequence elements enhance poly(A) site efficiency of the C2 complement gene and are phylogenetically conserved. , 1995, The EMBO journal.

[2]  B. Graveley,et al.  CPSF recognition of an HIV-1 mRNA 3'-processing enhancer: multiple sequence contacts involved in poly(A) site definition. , 1995, Genes & development.

[3]  M. Peterson.  Regulated immunoglobulin (Ig) RNA processing does not require specific cis-acting sequences: non-Ig RNA can be alternatively processed in B cells and plasma cells , 1994, Molecular and cellular biology.

[4]  C. Baker,et al.  Sequences homologous to 5' splice sites are required for the inhibitory activity of papillomavirus late 3' untranslated regions , 1994, Molecular and cellular biology.

[5]  Véronique Picard,et al.  A rapid and efficient one-tube PCR-based mutagenesis technique using Pfu DNA polymerase , 1994, Nucleic Acids Res..

[6]  Lung-Ji Chang,et al.  Presence of negative and positive cis-acting RNA splicing elements within and flanking the first tat coding exon of human immunodeficiency virus type 1 , 1994, Molecular and cellular biology.

[7]  J. Alwine,et al.  Direct interaction of the U1 snRNP-A protein with the upstream efficiency element of the SV40 late polyadenylation signal. , 1994, Genes & development.

[8]  W. Boelens,et al.  The human U1A snRNP protein regulates polyadenylation via a direct interaction with poly(A) polymerase , 1994, Cell.

[9]  L. Maquat,et al.  Upstream introns influence the efficiency of final intron removal and RNA 3'-end formation. , 1994, Genes & development.

[10]  N. Proudfoot,et al.  RNA 3' ends: formation and function--meeting review. , 1994, Genes & development.

[11]  T. Hohn,et al.  Pararetroviruses and retroviruses: a comparative review of viral structure and gene expression strategies. , 1994, Advances in virus research.

[12]  D. Purcell,et al.  Alternative splicing of human immunodeficiency virus type 1 mRNA modulates viral protein expression, replication, and infectivity , 1993, Journal of virology.

[13]  A. Sachs,et al.  Messenger RNA degradation in eukaryotes , 1993, Cell.

[14]  P. Sheridan,et al.  HIV-1 core promoter lacks a simple initiator element but contains a bipartite activator at the transcription start site. , 1993, The Journal of biological chemistry.

[15]  N. Proudfoot,et al.  Poly(A) signals and transcriptional pause sites combine to prevent interference between RNA polymerase II promoters. , 1993, The EMBO journal.

[16]  L. Maquat,et al.  Sequences within the last intron function in RNA 3'-end formation in cultured cells , 1993, Molecular and cellular biology.

[17]  J. Eggermont,et al.  Tat‐dependent occlusion of the HIV poly(A) site. , 1993, The EMBO journal.

[18]  J. Nevins,et al.  Alternative poly(A) site utilization during adenovirus infection coincides with a decrease in the activity of a poly(A) site processing factor , 1993, Molecular and cellular biology.

[19]  W. Boelens,et al.  The human U1 snRNP-Specific U1A protein inhibits polyadenylation of its own pre-mRNA , 1993, Cell.

[20]  M. Collin,et al.  A Rat CD4 Mutant Containing the gpl20-binding Site Mediates Human Immunodeficiency Virus Type 1 Infection By James H. M. Simon,~ Chamorro Somoza,* , 1993 .

[21]  A. Phelan,et al.  Herpes simplex virus IE63 acts at the posttranscriptional level to stimulate viral mRNA 3' processing , 1992, Journal of virology.

[22]  E. Fleming,et al.  Activation of HIV‐1 pre‐mRNA 3′ processing in vitro requires both an upstream element and TAR. , 1992, The EMBO journal.

[23]  J. Cherrington,et al.  Upstream sequences and cap proximity in the regulation of polyadenylation in ground squirrel hepatitis virus , 1992, Journal of virology.

[24]  M. Imperiale,et al.  Relative roles of signals upstream of AAUAAA and promoter proximity in regulation of human immunodeficiency virus type 1 mRNA 3' end formation , 1992, Molecular and cellular biology.

[25]  Susan M. Berget,et al.  Are vertebrate exons scanned during splice-site selection? , 1992, Nature.

[26]  J. Alwine,et al.  Elements upstream of the AAUAAA within the human immunodeficiency virus polyadenylation signal are required for efficient polyadenylation in vitro , 1992, Molecular and cellular biology.

[27]  H. Kung,et al.  Transcriptional interaction between retroviral long terminal repeats (LTRs): mechanism of 5' LTR suppression and 3' LTR promoter activation of c-myc in avian B-cell lymphomas , 1992, Journal of virology.

[28]  J. Cherrington,et al.  Regulation of polyadenylation in human immunodeficiency virus (HIV): contributions of promoter proximity and upstream sequences. , 1992, The EMBO journal.

[29]  R. Russnak.  Regulation of polyadenylation in hepatitis B viruses: stimulation by the upstream activating signal PS1 is orientation-dependent, distance-independent, and additive. , 1991, Nucleic acids research.

[30]  J. Taylor,et al.  Regulation of polyadenylation of hepatitis delta virus antigenomic RNA , 1991, Journal of virology.

[31]  C. Dabrowski,et al.  Simian virus 40 late mRNA leader sequences involved in augmenting mRNA accumulation via multiple mechanisms, including increased polyadenylation efficiency , 1991, Journal of virology.

[32]  W. Ellmeier,et al.  Mature mRNA 3′ end formation stimulates RNA export from the nucleus. , 1991, The EMBO journal.

[33]  C. Baker,et al.  An element in the bovine papillomavirus late 3' untranslated region reduces polyadenylated cytoplasmic RNA levels , 1991, Journal of virology.

[34]  B. Cullen,et al.  Efficient polyadenylation within the human immunodeficiency virus type 1 long terminal repeat requires flanking U3-specific sequences , 1991, Journal of virology.

[35]  J. Alwine,et al.  The human immunodeficiency virus type 1 polyadenylylation signal: a 3' long terminal repeat element upstream of the AAUAAA necessary for efficient polyadenylylation. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[36]  M. Imperiale,et al.  Involvement of long terminal repeat U3 sequences overlapping the transcription control region in human immunodeficiency virus type 1 mRNA 3' end formation , 1991, Molecular and cellular biology.

[37]  Joseph R. Nevins,et al.  The HTLV-I rex response element mediates a novel form of mRNA polyadenylation , 1991, Cell.

[38]  N. Proudfoot,et al.  Occlusion of the HIV poly(A) site. , 1991, Genes & development.

[39]  T. Hohn,et al.  A dissection of the cauliflower mosaic virus polyadenylation signal. , 1991, Genes & development.

[40]  H. Temin,et al.  The U3 region is not necessary for 3' end formation of spleen necrosis virus RNA , 1990, Journal of virology.

[41]  S. Rose,et al.  In vitro polyadenylation is stimulated by the presence of an upstream intron. , 1990, Genes & development.

[42]  R. Jackson,et al.  Do the poly(A) tail and 3′ untranslated region control mRNA translation? , 1990, Cell.

[43]  T. Hohn,et al.  Proximity to the promoter inhibits recognition of cauliflower mosaic virus polyadenylation signal , 1990, Nature.

[44]  D. Ganem,et al.  Sequences 5' to the polyadenylation signal mediate differential poly(A) site use in hepatitis B viruses. , 1990, Genes & development.

[45]  M. Mathews,et al.  HIV-1 Tat protein increases transcriptional initiation and stabilizes elongation , 1989, Cell.

[46]  J. Alwine,et al.  Efficiency of utilization of the simian virus 40 late polyadenylation site: effects of upstream sequences , 1989, Molecular and cellular biology.

[47]  N. Proudfoot,et al.  Definition of an efficient synthetic poly(A) site. , 1989, Genes & development.

[48]  Christopher W. J. Smith,et al.  Mutually exclusive splicing of α-tropomyosin exons enforced by an unusual lariat branch point location: Implications for constitutive splicing , 1989, Cell.

[49]  M. Braddock,et al.  Synthesis of a gene for the HIV transactivator protein TAT by a novel single stranded approach involving in vivo gap repair. , 1988, Nucleic acids research.

[50]  H. Temin,et al.  A promoterless retroviral vector indicates that there are sequences in U3 required for 3' RNA processing. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[51]  J. Coffin,et al.  Differential transcription from the long terminal repeats of integrated avian leukosis virus DNA , 1986, Journal of virology.

[52]  H. Gendelman,et al.  Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone , 1986, Journal of virology.

[53]  B. Cullen,et al.  The role of avian retroviral LTRs in the regulation of gene expression and viral replication. , 1985, Advances in virus research.

[54]  B. Cullen,et al.  Transcriptional interference in avian retroviruses—implications for the promoter insertion model of leukaemogenesis , 1984, Nature.

[55]  J. Vieira,et al.  The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. , 1982, Gene.