DOI: 10.1126/science.1064987. A Combined Experimental and Computational Strategy to Define Protein Interaction Networks for Peptide Recognition Modules

A Combined Experimental and Computational Strategy to Define Protein Interaction Networks for Peptide Recognition Modules

10.1126/science.1064987

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Peptide recognition modules mediate many protein-protein interactions critical for the assembly of macromolecular complexes. Complete genome sequences have revealed thousands of these domains, requiring improved methods for identifying their physiologically relevant binding partners. We have developed a strategy combining computational prediction of interactions from phage-display ligand consensus sequences with large-scale two-hybrid physical interaction tests. Application to yeast SH3 domains generated a phage-display network containing 394 interactions among 206 proteins and a two-hybrid network containing 233 interactions among 145 proteins. Graph theoretic analysis identified 59 highly likely interactions common to both networks. Las17 (Bee1), a member of the Wiskott-Aldrich Syndrome protein (WASP) family of actin-assembly proteins, showed multiple SH3 interactions, many of which were confirmed in vivo by coimmunoprecipitation.

Bibliography

Tong, A. H. Y., Drees, B., Nardelli, G., Bader, G. D., Brannetti, B., Castagnoli, L., Evangelista, M., Ferracuti, S., Nelson, B., Paoluzi, S., Quondam, M., Zucconi, A., Hogue, C. W. V., Fields, S., Boone, C., & Cesareni, G. (2002). A Combined Experimental and Computational Strategy to Define Protein Interaction Networks for Peptide Recognition Modules. Science, 295(5553), 321â324.

Authors 16

Amy Hin Yan Tong (first)
Becky Drees (additional)
Giuliano Nardelli (additional)
Gary D. Bader (additional)
Barbara Brannetti (additional)
Luisa Castagnoli (additional)
Marie Evangelista (additional)
Silvia Ferracuti (additional)
Bryce Nelson (additional)
Serena Paoluzi (additional)
Michele Quondam (additional)
Adriana Zucconi (additional)
Christopher W. V. Hogue (additional)
Stanley Fields (additional)
Charles Boone (additional)
Gianni Cesareni (additional)

References 29 Referenced 568

10.1126/science.278.5346.2075
10.1126/science.8438166
A. E. Salcini et al. Genes Dev. 11 2239 (1997). (10.1101/gad.11.17.2239)
M. F. Moran et al. Proc. Natl. Acad. Sci. U.S.A. 87 8622 (1990). (10.1073/pnas.87.21.8622)
S. Paoluzi et al. EMBO J. 17 6541 (1998). (10.1093/emboj/17.22.6541)
10.1093/nar/25.17.3389
The following supplementary material presented as a description figure or table is available on Science Online at www.sciencemag.org/cgi/content/full/1064987/DC1: supplementary fig. 1 (S1) an alignment of the yeast SH3 domains; supplementary note 1 (S2) materials and methods for the phage-display analysis; supplementary table 1 (S3) the selected phage-display peptides that bind to each SH3 domain; supplementary note 2 (S4) the method for creating the position-specific scoring matrix used to rank the yeast peptides containing ligand consensus sequences; supplementary table 2 (S5) the binding and nonbinding peptides for each SH3 domain; supplementary table 3 (S6) the ligands predicted by the profile analysis; supplementary note 3 (S7) the methods used for visualization of protein-protein interaction networks; supplementary fig. 2 (S8) the phage-display network with each node assigned a gene name; supplementary note 4 (S9) the two-hybrid materials and methods; supplementary table 4 (S10) the positive interactions derived from the two-hybrid assays; supplementary fig. 3 (S11) the two-hybrid interactions derived from directed assays of SH3 domains with proline-rich targets; supplementary fig. 4 (S12) the two-hybrid network with each node assigned a gene name; supplementary note 5 (S13) evidence to support the protein-protein interactions predicted in the overlap network; supplementary note 6 (S14) the materials and methods used for the Las17 coimmunoprecipitation experiments; supplementary note 7 (S15) the materials and methods used to construct the plasmids encoding Bzz1W531S (476-633) Bzz1W615S (476-633) and Bzz1W531S W615S (476-633); supplementary fig. 5 (S16) the cortical patchlike localization of Bzz1-GFP (green fluorescent protein) Bbc1-GFP Ysc84-GFP Ynl094w-GFP and the synthetic-lethal phenotype associated with bzz1 Δ vrp1 Δ double mutants; supplementary fig. 6 (S17) the ELISA assay results for phage-display analysis of the Las17 proline-rich peptides; supplementary table 5 (S18) the positive two-hybrid interactions and their BIND identification numbers.
References for all the genes mentioned in this study can be found at the following databases: Proteome (www.proteome.com/) Saccharomyces Genome Database () and the Munich Information Center for Protein Sequences (www.mips.biochem.mpg.de/). Yeast protein-protein interaction data sets can be found at Proteome and the Munich Information Center for Protein Sequences.
10.1242/jcs.114.7.1253
10.1038/82360
10.1093/nar/29.1.242
{'key': 'e_1_3_1_13_2', 'unstructured': ''}
10.1126/science.7624781
10.1091/mbc.8.2.367
10.1083/jcb.138.1.95
10.1016/S0960-9822(99)80218-8
M. Evangelista et al. J. Cell Biol. 148 353 (2000). (10.1083/jcb.148.2.353)
10.1093/genetics/152.3.881
A. Madania et al. Mol. Biol. Cell 10 3521 (1999). (10.1091/mbc.10.10.3521)
10.1038/35001009
T. Ito et al. Proc. Natl. Acad. Sci. U.S.A. 98 4569 (2001). (10.1073/pnas.061034498)
10.1083/jcb.200104057
The phage-display analysis should identify a subset of the natural interactions mediated directly by short peptides missing those that require either longer peptides or further stabilization by noncontiguous residues; moreover some of the ligands predicted by phage display may not be surface-exposed within a folded protein. In the case of the two-hybrid interactions there is a recognized potential for false-positives in part due to overexpression and nuclear targeting of the fusion proteins and because we are conducting screens of yeast proteins in yeast cells the interactions may not be direct. Further some of the SH3 domains screened by two-hybrid interactions were not included in the phage-display network and vice versa because some bait proteins yielded results only in one assay.
YPD (www.proteome.com/DB-demo/intro-to-YPD.html) contained 89 interactions that involved one of the SH3 domain proteins that we studied. Of these interactions 17 were found in the phage-display network (394 interactions in total) 16 in the two-hybrid network (233 interactions in total) and 13 in the overlap network (59 interactions in total). Thus the overlap network maintains most of the literature-validated interactions of the phage-display and two-hybrid network.
The SH3 domains of yeast type I myosins bind to a proline-rich ligand in Bbc1 (Mti1) (Fig. 3B) (27) and a BBC1 deletion bbc1 Δ suppresses the temperature sensitivity and endocytosis defects of vrp1 mutants (27).
We found that Bzz1-GFP concentrated in cortical patchlike structures (7) which localized to sites of polarized growth suggesting that Bzz1 may be targeted to cortical actin patches like components of the Las17-Vrp1-Myo3/5 complex (16 17). Moreover a similar patchlike localization pattern was observed for Ynl094w-GFP Bbc1-GFP and Ysc84-GFP (7) supporting the predicted Bzz1-Ynl094w Bbc1-Las17 Bbc1-Ynl094w and Ysc84-Las17 interactions (Fig. 3B). From a synthetic lethal screen in which a strain carrying a complete deletion of the BZZ1 gene MAT α bzz1 Δ was crossed to an ordered array of ∼2000 different MAT a gene-deletion strains (28) we found that bzz1 Δ vrp1 Δ double-mutant meiotic progeny showed reduced fitness specifically (7). This genetic interaction suggests that Bzz1 functionally associates with Bee1-Vrp1-Myo3/5 actin-assembly complex.
J. Mochida T. Yamamoto K. Fujimura-Kamada K. Tanaka personal communication.
10.1126/science.285.5429.901
We thank B. Kay for the PxxP biased library; A. Burgess K. Kennedy C. Lee N. Melo A. Parsons and I. Pot for help with the conventional two-hybrid screens; and A. Davidson for comments on the phage-display network. Supported by grants from the Canadian Institute of Health Research (CIHR) and the National Cancer Institute of Canada (C.B.); CIHR (C.H.); AIRC the European Union framework 4 program and the CNR target project Biotechnology (G.C.); and the National Center for Research Resources (P41 RR11823) (S.F.). C.B. is an International Research Scholar of the Howard Hughes Medical Institute and S.F. is an investigator of the Howard Hughes Medical Institute.

Dates

Type	When
Created	23 years ago (July 27, 2002, 5:47 a.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 5:48 p.m.)
Indexed	6 days, 15 hours ago (Aug. 19, 2025, 7:09 a.m.)
Issued	23 years, 7 months ago (Jan. 11, 2002)
Published	23 years, 7 months ago (Jan. 11, 2002)
Published Print	23 years, 7 months ago (Jan. 11, 2002)

Funders 0

None

BibTeX

@article{Tong_2002, title={A Combined Experimental and Computational Strategy to Define Protein Interaction Networks for Peptide Recognition Modules}, volume={295}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1064987}, DOI={10.1126/science.1064987}, number={5553}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Tong, Amy Hin Yan and Drees, Becky and Nardelli, Giuliano and Bader, Gary D. and Brannetti, Barbara and Castagnoli, Luisa and Evangelista, Marie and Ferracuti, Silvia and Nelson, Bryce and Paoluzi, Serena and Quondam, Michele and Zucconi, Adriana and Hogue, Christopher W. V. and Fields, Stanley and Boone, Charles and Cesareni, Gianni}, year={2002}, month=jan, pages={321–324} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        19
      ]
    ],
    "date-time": "2025-08-19T11:09:37Z",
    "timestamp": 1755601777652
  },
  "reference-count": 29,
  "publisher": "American Association for the Advancement of Science (AAAS)",
  "issue": "5553",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2002,
        1,
        11
      ]
    ]
  },
  "abstract": "<jats:p>Peptide recognition modules mediate many protein-protein interactions critical for the assembly of macromolecular complexes. Complete genome sequences have revealed thousands of these domains, requiring improved methods for identifying their physiologically relevant binding partners. We have developed a strategy combining computational prediction of interactions from phage-display ligand consensus sequences with large-scale two-hybrid physical interaction tests. Application to yeast SH3 domains generated a phage-display network containing 394 interactions among 206 proteins and a two-hybrid network containing 233 interactions among 145 proteins. Graph theoretic analysis identified 59 highly likely interactions common to both networks. Las17 (Bee1), a member of the Wiskott-Aldrich Syndrome protein (WASP) family of actin-assembly proteins, showed multiple SH3 interactions, many of which were confirmed in vivo by coimmunoprecipitation.</jats:p>",
  "DOI": "10.1126/science.1064987",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        27
      ]
    ],
    "date-time": "2002-07-27T09:47:15Z",
    "timestamp": 1027763235000
  },
  "page": "321-324",
  "source": "Crossref",
  "is-referenced-by-count": 568,
  "title": "A Combined Experimental and Computational Strategy to Define Protein Interaction Networks for Peptide Recognition Modules",
  "prefix": "10.1126",
  "volume": "295",
  "author": [
    {
      "given": "Amy Hin Yan",
      "family": "Tong",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Banting and Best Department of Medical Research and Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada M5G 1L6."
        }
      ]
    },
    {
      "given": "Becky",
      "family": "Drees",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Genome Sciences and Department of Medicine, University of Washington, Box 357730, Seattle, WA 98195, USA."
        }
      ]
    },
    {
      "given": "Giuliano",
      "family": "Nardelli",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Gary D.",
      "family": "Bader",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, Canada M5S 1A8 and Department of Biochemistry, University of Toronto, Ontario, Canada M5G 1L6."
        }
      ]
    },
    {
      "given": "Barbara",
      "family": "Brannetti",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Luisa",
      "family": "Castagnoli",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Marie",
      "family": "Evangelista",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, Queens University, Kingston, Ontario, Canada K7L 3N6."
        }
      ]
    },
    {
      "given": "Silvia",
      "family": "Ferracuti",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Bryce",
      "family": "Nelson",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, Queens University, Kingston, Ontario, Canada K7L 3N6."
        }
      ]
    },
    {
      "given": "Serena",
      "family": "Paoluzi",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Michele",
      "family": "Quondam",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Adriana",
      "family": "Zucconi",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    },
    {
      "given": "Christopher W. V.",
      "family": "Hogue",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, Canada M5S 1A8 and Department of Biochemistry, University of Toronto, Ontario, Canada M5G 1L6."
        }
      ]
    },
    {
      "given": "Stanley",
      "family": "Fields",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Genome Sciences and Department of Medicine, University of Washington, Box 357730, Seattle, WA 98195, USA."
        },
        {
          "name": "Howard Hughes Medical Institute, University of Washington, Box 353360, Seattle, WA 98195, USA."
        }
      ]
    },
    {
      "given": "Charles",
      "family": "Boone",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Banting and Best Department of Medical Research and Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada M5G 1L6."
        },
        {
          "name": "Department of Biology, Queens University, Kingston, Ontario, Canada K7L 3N6."
        }
      ]
    },
    {
      "given": "Gianni",
      "family": "Cesareni",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, 00133 Rome, Italy."
        }
      ]
    }
  ],
  "member": "221",
  "reference": [
    {
      "key": "e_1_3_1_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.278.5346.2075"
    },
    {
      "key": "e_1_3_1_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8438166"
    },
    {
      "key": "e_1_3_1_4_2",
      "doi-asserted-by": "crossref",
      "unstructured": "A. E. Salcini et al. Genes Dev. 11 2239 (1997).",
      "DOI": "10.1101/gad.11.17.2239"
    },
    {
      "key": "e_1_3_1_5_2",
      "doi-asserted-by": "crossref",
      "unstructured": "M. F. Moran et al. Proc. Natl. Acad. Sci. U.S.A. 87 8622 (1990).",
      "DOI": "10.1073/pnas.87.21.8622"
    },
    {
      "key": "e_1_3_1_6_2",
      "doi-asserted-by": "crossref",
      "unstructured": "S. Paoluzi et al. EMBO J. 17 6541 (1998).",
      "DOI": "10.1093/emboj/17.22.6541"
    },
    {
      "key": "e_1_3_1_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/25.17.3389"
    },
    {
      "key": "e_1_3_1_8_2",
      "unstructured": "The following supplementary material presented as a description figure or table is available on Science Online at www.sciencemag.org/cgi/content/full/1064987/DC1: supplementary fig. 1 (S1) an alignment of the yeast SH3 domains; supplementary note 1 (S2) materials and methods for the phage-display analysis; supplementary table 1 (S3) the selected phage-display peptides that bind to each SH3 domain; supplementary note 2 (S4) the method for creating the position-specific scoring matrix used to rank the yeast peptides containing ligand consensus sequences; supplementary table 2 (S5) the binding and nonbinding peptides for each SH3 domain; supplementary table 3 (S6) the ligands predicted by the profile analysis; supplementary note 3 (S7) the methods used for visualization of protein-protein interaction networks; supplementary fig. 2 (S8) the phage-display network with each node assigned a gene name; supplementary note 4 (S9) the two-hybrid materials and methods; supplementary table 4 (S10) the positive interactions derived from the two-hybrid assays; supplementary fig. 3 (S11) the two-hybrid interactions derived from directed assays of SH3 domains with proline-rich targets; supplementary fig. 4 (S12) the two-hybrid network with each node assigned a gene name; supplementary note 5 (S13) evidence to support the protein-protein interactions predicted in the overlap network; supplementary note 6 (S14) the materials and methods used for the Las17 coimmunoprecipitation experiments; supplementary note 7 (S15) the materials and methods used to construct the plasmids encoding Bzz1W531S (476-633) Bzz1W615S (476-633) and Bzz1W531S W615S (476-633); supplementary fig. 5 (S16) the cortical patchlike localization of Bzz1-GFP (green fluorescent protein) Bbc1-GFP Ysc84-GFP Ynl094w-GFP and the synthetic-lethal phenotype associated with bzz1 \u0394 vrp1 \u0394 double mutants; supplementary fig. 6 (S17) the ELISA assay results for phage-display analysis of the Las17 proline-rich peptides; supplementary table 5 (S18) the positive two-hybrid interactions and their BIND identification numbers."
    },
    {
      "key": "e_1_3_1_9_2",
      "unstructured": "References for all the genes mentioned in this study can be found at the following databases: Proteome (www.proteome.com/) Saccharomyces Genome Database () and the Munich Information Center for Protein Sequences (www.mips.biochem.mpg.de/). Yeast protein-protein interaction data sets can be found at Proteome and the Munich Information Center for Protein Sequences."
    },
    {
      "key": "e_1_3_1_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1242/jcs.114.7.1253"
    },
    {
      "key": "e_1_3_1_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/82360"
    },
    {
      "key": "e_1_3_1_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/29.1.242"
    },
    {
      "key": "e_1_3_1_13_2",
      "unstructured": ""
    },
    {
      "key": "e_1_3_1_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.7624781"
    },
    {
      "key": "e_1_3_1_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.8.2.367"
    },
    {
      "key": "e_1_3_1_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.138.1.95"
    },
    {
      "key": "e_1_3_1_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0960-9822(99)80218-8"
    },
    {
      "key": "e_1_3_1_18_2",
      "doi-asserted-by": "crossref",
      "unstructured": "M. Evangelista et al. J. Cell Biol. 148 353 (2000).",
      "DOI": "10.1083/jcb.148.2.353"
    },
    {
      "key": "e_1_3_1_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/genetics/152.3.881"
    },
    {
      "key": "e_1_3_1_20_2",
      "doi-asserted-by": "crossref",
      "unstructured": "A. Madania et al. Mol. Biol. Cell 10 3521 (1999).",
      "DOI": "10.1091/mbc.10.10.3521"
    },
    {
      "key": "e_1_3_1_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/35001009"
    },
    {
      "key": "e_1_3_1_22_2",
      "doi-asserted-by": "crossref",
      "unstructured": "T. Ito et al. Proc. Natl. Acad. Sci. U.S.A. 98 4569 (2001).",
      "DOI": "10.1073/pnas.061034498"
    },
    {
      "key": "e_1_3_1_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.200104057"
    },
    {
      "key": "e_1_3_1_24_2",
      "unstructured": "The phage-display analysis should identify a subset of the natural interactions mediated directly by short peptides missing those that require either longer peptides or further stabilization by noncontiguous residues; moreover some of the ligands predicted by phage display may not be surface-exposed within a folded protein. In the case of the two-hybrid interactions there is a recognized potential for false-positives in part due to overexpression and nuclear targeting of the fusion proteins and because we are conducting screens of yeast proteins in yeast cells the interactions may not be direct. Further some of the SH3 domains screened by two-hybrid interactions were not included in the phage-display network and vice versa because some bait proteins yielded results only in one assay."
    },
    {
      "key": "e_1_3_1_25_2",
      "unstructured": "YPD (www.proteome.com/DB-demo/intro-to-YPD.html) contained 89 interactions that involved one of the SH3 domain proteins that we studied. Of these interactions 17 were found in the phage-display network (394 interactions in total) 16 in the two-hybrid network (233 interactions in total) and 13 in the overlap network (59 interactions in total). Thus the overlap network maintains most of the literature-validated interactions of the phage-display and two-hybrid network."
    },
    {
      "key": "e_1_3_1_26_2",
      "unstructured": "The SH3 domains of yeast type I myosins bind to a proline-rich ligand in Bbc1 (Mti1) (Fig. 3B) (27) and a BBC1 deletion bbc1 \u0394 suppresses the temperature sensitivity and endocytosis defects of vrp1 mutants (27)."
    },
    {
      "key": "e_1_3_1_27_2",
      "unstructured": "We found that Bzz1-GFP concentrated in cortical patchlike structures (7) which localized to sites of polarized growth suggesting that Bzz1 may be targeted to cortical actin patches like components of the Las17-Vrp1-Myo3/5 complex (16 17). Moreover a similar patchlike localization pattern was observed for Ynl094w-GFP Bbc1-GFP and Ysc84-GFP (7) supporting the predicted Bzz1-Ynl094w Bbc1-Las17 Bbc1-Ynl094w and Ysc84-Las17 interactions (Fig. 3B). From a synthetic lethal screen in which a strain carrying a complete deletion of the BZZ1 gene MAT \u03b1 bzz1 \u0394 was crossed to an ordered array of \u223c2000 different MAT a gene-deletion strains (28) we found that bzz1 \u0394 vrp1 \u0394 double-mutant meiotic progeny showed reduced fitness specifically (7). This genetic interaction suggests that Bzz1 functionally associates with Bee1-Vrp1-Myo3/5 actin-assembly complex."
    },
    {
      "key": "e_1_3_1_28_2",
      "unstructured": "J. Mochida T. Yamamoto K. Fujimura-Kamada K. Tanaka personal communication."
    },
    {
      "key": "e_1_3_1_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.285.5429.901"
    },
    {
      "key": "e_1_3_1_30_2",
      "unstructured": "We thank B. Kay for the PxxP biased library; A. Burgess K. Kennedy C. Lee N. Melo A. Parsons and I. Pot for help with the conventional two-hybrid screens; and A. Davidson for comments on the phage-display network. Supported by grants from the Canadian Institute of Health Research (CIHR) and the National Cancer Institute of Canada (C.B.); CIHR (C.H.); AIRC the European Union framework 4 program and the CNR target project Biotechnology (G.C.); and the National Center for Research Resources (P41 RR11823) (S.F.). C.B. is an International Research Scholar of the Howard Hughes Medical Institute and S.F. is an investigator of the Howard Hughes Medical Institute."
    }
  ],
  "container-title": "Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://www.science.org/doi/pdf/10.1126/science.1064987",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2024,
        1,
        9
      ]
    ],
    "date-time": "2024-01-09T22:48:59Z",
    "timestamp": 1704840539000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.science.org/doi/10.1126/science.1064987"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2002,
        1,
        11
      ]
    ]
  },
  "references-count": 29,
  "journal-issue": {
    "issue": "5553",
    "published-print": {
      "date-parts": [
        [
          2002,
          1,
          11
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1126/science.1064987"
  ],
  "URL": "http://dx.doi.org/10.1126/science.1064987",
  "relation": {
    "has-review": [
      {
        "id-type": "doi",
        "id": "10.3410/f.1003682.48205",
        "asserted-by": "object"
      },
      {
        "id-type": "doi",
        "id": "10.3410/f.1003682.38006",
        "asserted-by": "object"
      },
      {
        "id-type": "doi",
        "id": "10.3410/f.1003682.207563",
        "asserted-by": "object"
      }
    ]
  },
  "ISSN": [
    "0036-8075",
    "1095-9203"
  ],
  "subject": [],
  "container-title-short": "Science",
  "published": {
    "date-parts": [
      [
        2002,
        1,
        11
      ]
    ]
  }
}