p10 Single-stranded nucleic acid binding protein from murine leukemia virus binds metal ions via the peptide sequence Cys26-X2-Cys29-X4-His34-X4-Cys39

Roberts, William J.; Pan, Tao; Elliott, James I.; Coleman, Joseph E.; Williams, Kenneth R.

doi:10.1021/bi00452a024

Cited by 39 publications

(37 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This peptide serves as an important model system, as the bound zinc atom is known to exist in a 3Cys-1His tetrahedral environment (South et al, 1989;Summers et al, 1990). Several other peptides and proteins that contain the CCHC motif have been shown by UV-visible and NMR spectroscopic methods to bind zinc via 3Cys and 1 His residues, forming a tetrahedral metal coordination site (Green & Berg, 1989Roberts et al, 1989;Fitzgerald & Coleman, 1991;South et al, 1991). Although nonlinear least-squares fitting of the Fourier-filtered (first coordination sphere) EXAFS data obtained for HIV1-Fl afforded several solutions that are consistent with the EXAFS data alone (Chance, M., pers.…”

Section: Exafs Spectra Of Intact Retroviruses and The Relevance Of CCmentioning

confidence: 99%

“…Without exception, retroviral NC proteins (and their gag precursors) contain one or two copies of an invariant CCHC array, Cys-X,-Cys-X,-His-X4-Cys (X = variable or conservatively substituted amino acid) (Henderson et al, 1981), that are essential for RNA genome packaging and infectivity (Gorelick et al, 1988(Gorelick et al, , 1990Meric & Goff, 1989;Aldovini & Young, 1990) and have been proposed to function as zinc-binding domains (Berg, 1986). Although the arrays are capable of binding zinc in vitro (Green & Berg, 1989Roberts et al, 1989;South et al, 1989South et al, , 1990aSouth et al, ,b, 1991Summers et al, 1990;Fitzgerald & Coleman, 1991;Omichinski et al, 1991), the physiological relevance of this CCHC zinc finger motif (previously referred to as a retroviral-type or zinc finger-like motif) has not been firmly established. In fact, it has been reported that avian myeloblastosis virus (AMV) particles lack significant amounts of zinc (Jentoft et al, 1988), which has led to the suggestion that the conserved arrays do not function as zinc binding fingers in mature viruses (Jentoft et al, 1988;Katz & Jentoft, 1989).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution‐state structure of the nucleocapsid protein from HIV‐1

Henderson²,

et al. 1992

View full text Add to dashboard Cite

All retroviral nucleocapsid (NC) proteins contain one or two copies of an invariant 3Cys‐1His array (CCHC = C‐X2‐C‐X4‐H‐X4‐C; C = Cys, H = His, X = variable amino acid) that are essential for RNA genome packaging and infectivity and have been proposed to function as zinc‐binding domains. Although the arrays are capable of binding zinc in vitro, the physiological relevance of zinc coordination has not been firmly established. We have obtained zinc‐edge extended X‐ray absorption fine structure (EXAFS) spectra for intact retroviruses in order to determine if virus‐bound zinc, which is present in quantities nearly stoichiometric with the CCHC arrays (Bess, J.W., Jr., Powell, P.J., Issaq, H.J., Schumack, L.J., Grimes, M.K., Henderson, L.E., & Arthur, L.O., 1992, J. Virol. 66, 840–847), exists in a unique coordination environment. The viral EXAFS spectra obtained are remarkably similar to the spectrum of a model CCHC zinc finger peptide with known 3Cys‐1His zinc coordination structure. This finding, combined with other biochemical results, indicates that the majority of the viral zinc is coordinated to the NC CCHC arrays in mature retroviruses. Based on these findings, we have extended our NMR studies of the HIV‐1 NC protein and have determined its three‐dimensional solution‐state structure. The CCHC arrays of HIV‐1 NC exist as independently folded, noninteracting domains on a flexible polypeptide chain, with conservatively substituted aromatic residues forming hydrophobic patches on the zinc finger surfaces. These residues are essential for RNA genome recognition, and fluorescence measurements indicate that at least one residue (Trp37) participates directly in binding to nucleic acids in vitro. The NC is only the third HIV‐1 protein to be structurally characterized, and the combined EXAFS, structural, and nucleic acid‐binding results provide a basis for the rational design of new NC‐targeted antiviral agents and vaccines for the control of AIDS.

show abstract

Section: Exafs Spectra Of Intact Retroviruses and The Relevance Of CCmentioning

confidence: 99%

mentioning

confidence: 99%

Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution‐state structure of the nucleocapsid protein from HIV‐1

Henderson²,

et al. 1992

View full text Add to dashboard Cite

show abstract

“…An examination of the predicted SLU7 protein sequence revealed a region that bears striking similarity to a cysteine-rich zinc knuckle motif of retroviral nucleocapsid proteins {Table 2; Berg 1986;South and Summers 1990). The hallmark of this motif is the consensus sequence CX~CX4HX4C, which coordinates zinc in a manner reminiscent of classical zinc finger proteins (Schiff et al 1988;Roberts et al 1989;Green and Berg 1990;Summers et al 1990;Fitzgerald and Coleman 1991). Note that the relative spacing of cysteine and histidine residues is strictly specified in the case of nucleocapsid proteins, unlike the case of zinc finger proteins.…”

Section: Cloning the Slu7 Genementioning

confidence: 99%

An essential splicing factor, SLU7, mediates 3' splice site choice in yeast.

Frank¹,

Guthrie²

1992

Genes Dev.

141

103

View full text Add to dashboard Cite

Recently, we have reported the identification of several genes that exhibit genetic interactions with the U5 snRNA. Two of these genes, SL U4 and SL U7 (SL U: synergistic lethal with _ U5 snRNA), encode products required for the second catalytic step of splicing. To analyze the specific roles of SLU4 and SLU7, we have determined how mutants influence the relative usage of competing 3' splice sites. We find that mutations in SLU7 eliminate the normal 20-fold preference for 3' splice sites located >22 nucleotides downstream of the branchpoint. In contrast, mutations in SLU4 inhibit usage of all 3' splice sites, regardless of their location. This suggests that SLU7 is involved in the process of 3' splice site choice, whereas SLU4 fulfills a generic requirement for the second step. We show that SLU7 is an essential gene that contains a small motif with striking similarity to the cysteine-rich zinc knuckle of retroviral nucleocapsid proteins, which has been implicated in RNA binding. Mutational analysis of SL U7 indicates that this motif influences the efficiency, but not the sequence specificity, of 3' splice site selection. The identification of a component of the constitutive splicing machinery that can promote 3' splice site choice has potentially important implications for alternative splicing.[Key Words: Pre-mRNA splicing; Saccharomyces cerevisiae; SLU mutants; U5 snRNP; alternative splicing] Received July 6, 1992; revised version accepted August 27, 1992.Nuclear pre-mRNA introns are defined by three conserved sequence elements. Two mark the 5' and 3' exonintron boundaries while a third, the branchpoint sequence, is located just upstream of the 3' splice site. Unlike autocatalytic introns, these intron sequences are not sufficient to catalyze their own removal (Cech and Bass 1986). Instead, numerous cofactors, including the small nuclear ribonucleoprotein (snRNP) particles, are essential for splicing. These factors bind to the premRNA in a defined hierarchy that culminates in the assembly of a splicing-competent structure termed the spliceosome. Once this complex is assembled, intron removal proceeds by two consecutive catalytic steps. The first step entails a transesterification reaction between the 5' exon-intron junction and an adenosine residue within the branchpoint sequence that results in cleavage of the 5' exon from the intron. During the second step, cleavage occurs at the intron-3' exon border and the free 5' exon is concomitantly ligated to the 3' exon (Padgett et al. 1986;Guthrie 1991).Given the complex matrix of interactions that occur between the splicing factors and the pre-mRNA, a key problem in understanding the splicing process has been to determine which of these factors are responsible for the primary recognition of the pre-mRNA. The U1 and U2 snRNPs are important determinants of 5' splice site and branchpoint recognition (respectively) that function 1Corresponding author.by direct base-pairing between snRNA and pre-mRNA (for review, see Steitz et al. 1988). Unambiguous identification of the...

show abstract

“…Subsequent to assembly and budding, the gag poly (Berg, 1986) that the arrays function by coordinating zinc (Green & Berg, 1989Roberts et al, 1989;South et al, 1989South et al, , 1990a South et al, ,b, 1991Summers et al, 1990Summers et al, , 1992Fitzgerald & Coleman, 1991;Omichinski et al, 1991). In particular, recent zinc-edge extended X-ray absorption fine structure (EXAFS) measurements on intact retroviruses reveal that tightly associated viral zinc, which is present in quantities sufficient to populate the arrays , is coordinated t o the CCHC zinc fingers of the NC proteins in mature virions Summers et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

Zinc‐ and sequence‐dependent binding to nucleic acids by the N‐terminal zinc finger of the HIV‐1 nucleocapsid protein: NMR structure of the complex with the Psi‐site analog, dACGCC

1993

View full text Add to dashboard Cite

The nucleic acid interactive properties of a synthetic peptide with sequence of the N-terminal CCHC zinc finger (CCHC = Cys-X,-Cys-X4-His-X,-Cys; X = variable amino acid) of the human immunodeficiency virus (HIV) nucleocapsid protein, Zn(HIV1-Fl), have been studied by 'H NMR spectroscopy. Titration of Zn(HIV1-Fl) with oligodeoxyribonucleic acids containing different nucleotide sequences reveals, for the first time, sequencedependent binding that requires the presence of at least one guanosine residue for tight complex formation. The dynamics of complex formation are sensitive to the nature of the residues adjacent to guanosine, with residues on the 3' side of guanosine having the largest influence. An oligodeoxyribonucleotide with sequence corresponding to a portion of the HIV-1 psi-packaging signal, d(ACGCC), forms a relatively tight complex with Zn(HIV1-F1) (Kd = 5 x M). Two-dimensional nuclear Overhauser effect (NOESY) data indicate that the bound nucleic acid exists predominantly in a single-stranded, A-helical conformation, and the presence of more than a dozen intermolecular NOE cross peaks enabled three-dimensional modeling of the complex. The nucleic acid binds within a hydrophobic cleft on the peptide surface. This hydrophobic cleft is defined by the side chains of residues Val', Phe4, He1,, and AlaI3. Backbone amide protons of Phe4 and AlaI3 and the backbone carbonyl oxygen of Lys' that lie within this cleft appear to form hydrogen bonds with the guanosine 0 6 and NlH atoms, respectively. In addition, the positively charged side chain of Arg14 is ideally positioned for electrostatic interactions with the phosphodiester backbone of the nucleic acid. The structural findings provide a rationalization for the general conservation of these hydrophobic and basic residues in CCHC zinc fingers, and are consistent with site-directed mutagenesis results that implicate these residues as direct participants in viral genome recognition.Keywords: human immunodeficiency virus; NMR; nucleocapsid protein; viral genome recognition; zinc finger All retroviruses, including human immunodeficiency virus (HIV), encode a gag precursor polyprotein that functions in the recognition of viral RNA and in the assembly of virus particles (Bolognesi et al., 1978;Dickson et al., 1985). Subsequent to assembly and budding, the gag poly (Berg, 1986) that the arrays function by coordinating zinc (Green & Berg, 1989Roberts et al., 1989;South et al., 1989South et al., , 1990a South et al., ,b, 1991Summers et al., 1990Summers et al., , 1992Fitzgerald & Coleman, 1991;Omichinski et al., 1991). In particular, recent zinc-edge extended X-ray absorption fine structure (EXAFS) measurements on intact retroviruses reveal that tightly associated viral zinc, which is present in quantities sufficient to populate the arrays , is coordinated t o the CCHC zinc fingers of the NC proteins in mature virions Summers et al., 1992). The three-dimensional structures of the HIVl-NC protein and its constituent zinc finger domains have been determined to...

show abstract

p10 Single-stranded nucleic acid binding protein from murine leukemia virus binds metal ions via the peptide sequence Cys26-X2-Cys29-X4-His34-X4-Cys39

Cited by 39 publications

References 20 publications

Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution‐state structure of the nucleocapsid protein from HIV‐1

Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution‐state structure of the nucleocapsid protein from HIV‐1

An essential splicing factor, SLU7, mediates 3' splice site choice in yeast.

Zinc‐ and sequence‐dependent binding to nucleic acids by the N‐terminal zinc finger of the HIV‐1 nucleocapsid protein: NMR structure of the complex with the Psi‐site analog, dACGCC

Contact Info

Product

Resources

About