FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Virol / The predicted metal-binding region of the arterivirus helica...
Journal of virology2000; 74(11); 5213-5223; doi: 10.1128/jvi.74.11.5213-5223.2000

The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis.

Abstract: Equine arteritis virus (EAV), the prototype Arterivirus, is a positive-stranded RNA virus that expresses its replicase in the form of two large polyproteins of 1,727 and 3,175 amino acids. The functional replicase subunits (nonstructural proteins), which drive EAV genome replication and subgenomic mRNA transcription, are generated by extensive proteolytic processing. Subgenomic mRNA transcription involves an unusual discontinuous step and generates the mRNAs for structural protein expression. Previously, the phenotype of mutant EAV030F, which carries a single replicase point mutation (Ser-2429-->Pro), had implicated the nsp10 replicase subunit (51 kDa) in viral RNA synthesis, and in particular in subgenomic mRNA transcription. nsp10 contains an N-terminal (putative) metal-binding domain (MBD), located just upstream of the Ser-2429-->Pro mutation, and a helicase activity in its C-terminal part. We have now analyzed the N-terminal domain of nsp10 in considerable detail. A total of 38 mutants, most of them carrying specific single point mutations, were tested in the context of an EAV infectious cDNA clone. Variable effects on viral genome replication and subgenomic mRNA transcription were observed. In general, our results indicated that the MBD region, and in particular a set of 13 conserved Cys and His residues that are assumed to be involved in zinc binding, is essential for viral RNA synthesis. On the basis of these data and comparative sequence analyses, we postulate that the MBD may employ a rather unusual mode of zinc binding that could result in the association of up to four zinc cations with this domain. The region containing residue Ser-2429 may play the role of "hinge spacer," which connects the MBD to the rest of nsp10. Several mutations in this region specifically affected subgenomic mRNA synthesis. Furthermore, one of the MBD mutants was replication and transcription competent but did not produce infectious progeny virus. This suggests that nsp10 is involved in an as yet unidentified step of virion biogenesis.
Get Full Text
Publication Date: 2000-05-09 PubMed ID: 10799597PubMed Central: PMC110875DOI: 10.1128/jvi.74.11.5213-5223.2000Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research paper investigates the role of a particular region in the arterivirus helicase protein in creating a subgenomic mRNA, replicating the genome, and developing virions (virus particles) using the equine arteritis virus as its subject.

Background

  • The mentioned research is focused on the Equine arteritis virus (EAV), positioned as a prototype Arterivirus. This virus carries its replicase in two large proteins of 1,727 and 3,175 amino acids.
  • The functional subunits enabling EAV genome replication and subgenomic mRNA (messenger Ribonucleic Acid) transcription are a product of proteolytic processing.
  • The paper acknowledges previous analysis and findings concerning EAV030F, a mutant EAV featuring a single point mutation (Ser-2429–>Pro) which emphasized the role of the nsp10 replicase subunit in viral RNA (Ribonucleic Acid) synthesis and subgenomic mRNA transcription.

Exploration of nsp10

  • The researchers have conducted a detailed study of the N-terminal domain of nsp10. The domain contains an assumed metal-binding domain (MBD) upstream of the Ser-2429–>Pro mutation while its C-terminal holds helicase activity.
  • A total of 38 mutants—predominantly single point mutations—were tested in coherence with an EAV infectious cDNA clone.
  • A wide range of influence was observed on viral genome replication and subgenomic mRNA transcription due to these mutations.
  • The results indicated the crucial role of the MBD region, specifically a group of 13 conserved cysteine and histidine residues, on viral RNA synthesis, presumed to enable zinc binding.

Role of MBD and Ser-2429 region

  • The researchers have speculated that this MBD employs a unique method of zinc binding which could lead to an association of up to four zinc cations with this domain.
  • The residue containing Ser-2429 region possibly acts as a ‘hinge spacer’, linking the MBD to the rest of nsp10.
  • The research found that mutations in this region specifically affected subgenomic mRNA synthesis.
  • Interestingly, one of their MBD mutants could replicate and transcribe but was unable to produce infectious progeny virus. This finding pointed towards the possible involvement of nsp10 in yet-to-be-identified stages of virion (actual virus particle) biogenesis.

Cite This Article

APA
van Dinten LC, van Tol H, Gorbalenya AE, Snijder EJ. (2000). The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis. J Virol, 74(11), 5213-5223. https://doi.org/10.1128/jvi.74.11.5213-5223.2000

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 74
Issue: 11
Pages: 5213-5223

Researcher Affiliations

van Dinten, L C
  • Department of Virology, Center for Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
van Tol, H
    Gorbalenya, A E
      Snijder, E J

        MeSH Terms

        • Amino Acid Sequence
        • Animals
        • Cell Line
        • Cricetinae
        • Equartevirus / enzymology
        • Equartevirus / genetics
        • Equartevirus / physiology
        • Genetic Complementation Test
        • Genome, Viral
        • Metals / metabolism
        • Molecular Sequence Data
        • RNA Helicases / genetics
        • RNA Helicases / metabolism
        • RNA, Messenger / biosynthesis
        • RNA, Viral / biosynthesis
        • Virion / physiology

        Grant Funding

        • N01-CO-56000 / NCI NIH HHS

        References

        This article includes 73 references
        1. Barlow P N, Luisi B, Milner A, Elliot M, Everett R. Structure of the C3HC4 domain by 1H-nuclear magnetic resonance spectroscopy: a new structural class of zinc-finger.. J Mol Biol 1994;237:201–211.
          pubmed: 8126734
        2. Bass S H, Mulkerrin M G, Wells J A. A systematic mutational analysis of hormone-binding determinants in the human growth hormone receptor.. Proc Natl Acad Sci USA 1991;88:4498–4502.
          pmc: PMC51688pubmed: 2034689
        3. Bellon S F, Rodgers K K, Schatz D G, Coleman J E, Steitz T A. Crystal structure of the RAG1 dimerization domain reveals multiple zinc-binding motifs including a novel zinc binuclear cluster.. Nat Struct Biol 1997;4:586–591.
          pubmed: 9228952
        4. Berg J M, Godwin H A. Lessons from zinc-binding peptides.. Annu Rev Biophys Biomol Struct 1997;26:357–371.
          pubmed: 9241423
        5. Berg J M, Shi Y. The galvanization of biology: a growing appreciation for the roles of zinc.. Science 1996;271:1081–1085.
          pubmed: 8599083
        6. Borden K L B, Boddy M N, Lally J, O'Reilly N J, Martin S, Howe K, Solomon E, Freemont P S. The solution structure of the RING finger domain from the acute promyelocytic leukaemia proto-oncoprotein PML.. EMBO J 1995;14:1532–1541.
          pmc: PMC398240pubmed: 7729428
        7. Cavanagh D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae.. Arch Virol 1997;142:629–633.
          pubmed: 9349308
        8. Dannull J, Surovoy A, Jung G, Moelling K. Specific binding of HIV-1 nucleocapsid protein to PSI RNA in vitro requires N-terminal zinc finger and flanking basic amino acid residues.. EMBO J 1994;13:1525–1533.
          pmc: PMC394981pubmed: 8156990
        9. den Boon J A, Kleijnen M F, Spaan W J M, Snijder E J. Equine arteritis virus subgenomic mRNA synthesis: analysis of leader-body junctions and replicative-form RNAs.. J Virol 1996;70:4291–4298.
          pmc: PMC190361pubmed: 8676451
        10. den Boon J A, Snijder E J, Chirnside E D, de Vries A A F, Horzinek M C, Spaan W J M. Equine arteritis virus is not a togavirus but belongs to the coronaviruslike superfamily.. J Virol 1991;65:2910–2920.
          pmc: PMC240924pubmed: 1851863
        11. den Boon J A, Spaan W J M, Snijder E J. Equine arteritis virus subgenomic RNA transcription: UV inactivation and translation inhibition studies.. Virology 1995;213:364–372.
          pmc: PMC7131190pubmed: 7491761
        12. Denison M R, Spaan W J M, van der Meer Y, Gibson C A, Sims A C, Prentice E, Lu X T. The putative helicase of the coronavirus mouse hepatitis virus is processed from the replicase gene polyprotein and localizes in complexes that are active in viral RNA synthesis.. J Virol 1999;73:6862–6871.
          pmc: PMC112771pubmed: 10400784
        13. de Vries A A F, Chirnside E D, Bredenbeek P J, Gravestein L A, Horzinek M C, Spaan W J M. All subgenomic mRNAs of equine arteritis virus contain a common leader sequence.. Nucleic Acids Res 1990;18:3241–3247.
          pmc: PMC330929pubmed: 2162519
        14. de Vries A A F, Chirnside E D, Horzinek M C, Rottier P J M. Structural proteins of equine arteritis virus.. J Virol 1992;66:6294–6303.
          pmc: PMC240121pubmed: 1328669
        15. de Vries A A F, Horzinek M C, Rottier P J M, de Groot R J. The genome organization of the Nidovirales: similarities and differences between arteri-, toro-, and coronaviruses.. Semin Virol 1997;8:33–47.
          pmc: PMC7128191pubmed: 32288441
        16. Diamond S E, Kirkegaard K. Clustered charged-to-alanine mutagenesis of poliovirus RNA-dependent RNA polymerase yields multiple temperature-sensitive mutants defective in RNA synthesis.. J Virol 1994;68:863–876.
          pmc: PMC236523pubmed: 8289389
        17. Doll E R, Bryans J T, McCollum W H, Crowe M E W. Isolation of a filterable agent causing arteritis of horses and abortion by mares. Its differentiation from the equine abortion (influenza) virus.. Cornell Vet 1957;47:3–41.
          pubmed: 13397177
        18. Dracheva S, Koonin E V, Crute J J. Identification of the primase active site of the herpes simplex virus type 1 helicase-primase.. J Biol Chem 1995;270:14148–14153.
          pubmed: 7775476
        19. Freemont P S. The RING finger: a novel protein sequence motif related to the zinc finger.. Ann N Y Acad Sci 1993;684:174–192.
          pubmed: 8317827
        20. Glaser A L, de Vries A A F, Dubovi E J. Comparison of equine arteritis virus isolates using neutralizing monoclonal antibodies and identification of sequence changes in GL associated with neutralization resistance.. J Gen Virol 1995;76:2223–2233.
          pubmed: 7561759
        21. Godeny E K, Chen L, Kumar S N, Methven S L, Koonin E V, Brinton M A. Complete genomic sequence and phylogenetic analysis of the lactate dehydrogenase-elevating virus (LDV). Virology 1993;194:585–596.
          pmc: PMC7173116pubmed: 8389075
        22. Gorbalenya A E, Koonin E V. Comparative analysis of the amino acid sequences of the key enzymes of the replication and expression of positive-strand RNA viruses. Validity of the approach and functional and evolutionary implications.. Sov Sci Rev Sect D 1993;11:1–84.
        23. Gorbalenya A E, Koonin E V, Donchenko A P, Blinov V M. Coronavirus genome: prediction of putative functional domains in the non-structural polyprotein by comparative amino acid sequence analysis.. Nucleic Acids Res 1989;17:4847–4861.
          pmc: PMC318036pubmed: 2526320
        24. Gorelick R J, Chabot D J, Ott D E, Gagliardi T D, Rein A, Henderson L E, Arthur L O. Genetic analysis of the zinc finger in the Moloney murine leukemia virus nucleocapsid domain: replacement of zinc-coordinating residues with other zinc-coordinating residues yields noninfectious particles containing genomic RNA.. J Virol 1996;70:2593–2597.
          pmc: PMC190107pubmed: 8642691
        25. Gorelick R J, Fu W, Gagliardi T D, Bosche W J, Rein A, Henderson L E, Arthur L O. Characterization of the block in replication of nucleocapsid protein zinc finger mutants from Moloney murine leukemia virus.. J Virol 1999;73:8185–8195.
          pmc: PMC112836pubmed: 10482569
        26. Hassett D E, Condit R C. Targeted construction of temperature-sensitive mutations in vaccinia virus by replacing clustered charged residues with alanine.. Proc Natl Acad Sci USA 1994;91:4554–4558.
          pmc: PMC43824pubmed: 8183946
        27. Hayward-Lester A, Hewetson A, Beale E G, Oefner P J, Doris P A, Chilton B S. Cloning, characterization, and steroid-dependent posttranscriptional processing of RUSH-1 alpha and beta, two uteroglobin promoter-binding proteins.. Mol Endocrinol 1996;10:1335–1349.
          pubmed: 8923460
        28. Heusipp G, Harms U, Siddell S G, Ziebuhr J. Identification of an ATPase activity associated with a 71-kilodalton polypeptide encoded in gene 1 of the human coronavirus 229E.. J Virol 1997;71:5631–5634.
          pmc: PMC191807pubmed: 9188639
        29. Jang S K, Krausslich H G, Nicklin M J, Duke G M, Palmenberg A C, Wimmer E. A segment of the 5′ nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation.. J Virol 1988;62:2636–2643.
          pmc: PMC253694pubmed: 2839690
        30. Johnson R E, Henderson S T, Petes T D, Prakash S, Bankmann M, Prakash L. Saccharomyces cerevisiae RAD5-encoded DNA repair protein contains DNA helicase and zinc-binding sequence motifs and affects the stability of simple repetitive sequences in the genome.. Mol Cell Biol 1992;12:3807–3818.
          pmc: PMC360249pubmed: 1324406
        31. Khromykh A A, Westaway E G. Subgenomic replicons of the flavivirus Kunjin: construction and applications.. J Virol 1997;71:1497–1505.
          pmc: PMC191206pubmed: 8995675
        32. Kuger P, Godecke A, Breunig K D. A mutation in the Zn-finger of the GAL4 homolog LAC9 results in glucose repression of its target genes.. Nucleic Acids Res 1990;18:745–751.
          pmc: PMC330322pubmed: 2107531
        33. Kusakabe T, Richardson C C. The role of the zinc motif in sequence recognition by DNA primases.. J Biol Chem 1996;271:19563–19570.
          pubmed: 8702650
        34. Lai M M C, Baric R S, Brayton P R, Stohlman S A. Characterization of leader RNA sequences on the virion and mRNAs of mouse hepatitis virus, a cytoplasmic RNA virus.. Proc Natl Acad Sci USA 1984;81:3626–3630.
          pmc: PMC345271pubmed: 6328522
        35. Lai M M C, Cavanagh D. The molecular biology of coronaviruses.. Adv Virus Res 1997;48:1–100.
          pmc: PMC7130985pubmed: 9233431
        36. Landt O, Grunert H-P, Hahn U. A general method for rapid site-directed mutagenesis using the polymerase chain reaction.. Gene 1990;96:125–128.
          pubmed: 2265750
        37. Lium E K, Silverstein S. Mutational analysis of the herpes simplex virus type 1 ICP0 C3HC4 zinc ring finger reveals a requirement for ICP0 in the expression of the essential α27 gene.. J Virol 1997;71:8602–8614.
          pmc: PMC192324pubmed: 9343218
        38. Loeber G, Stenger J E, Ray S, Parsons R E, Anderson M E, Tegtmeyer P. The zinc finger region of simian virus 40 large T antigen is needed for hexamer assembly and origin melting.. J Virol 1991;65:3167–3174.
          pmc: PMC240973pubmed: 1851875
        39. MacLachlan N J, Balasuriya U B, Hedges J F, Schweidler T M, McCollum W H, Timoney P J, Hullinger P J, Patton J F. Serologic response of horses to the structural proteins of equine arteritis virus.. J Vet Diagn Investig 1998;10:229–236.
          pubmed: 9683071
        40. Mannhaupt G, Stucka R, Ehnle S, Vetter I, Feldmann H. Molecular analysis of yeast chromosome II between CMD1 and LYS2: the excision repair gene RAD16 located in this region belongs to a novel group of double-finger proteins.. Yeast 1992;8:397–408.
          pubmed: 1626431
        41. Marmorstein R, Carey M, Ptashne M, Harrison S C. DNA recognition by GAL4: structure of a protein-DNA complex.. Nature 1992;356:379–380.
          pubmed: 1557122
        42. Meulenberg J J M, de Meijer E J, Moormann R J M. Subgenomic RNAs of Lelystad virus contain a conserved leader-body junction sequence.. J Gen Virol 1993;74:1697–1701.
          pubmed: 8345361
        43. Meulenberg J J M, Hulst M M, de Meijer E J, Moonen P L J M, den Besten A, de Kluyver E P, Wensvoort G, Moormann R J M. Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV.. Virology 1993;192:62–72.
          pmc: PMC7173055pubmed: 8517032
        44. Molenkamp R, Rozier B C D, Greve S, Spaan W J M, Snijder E J. Isolation and characterization of an arterivirus defective interfering RNA genome.. J Virol 2000;74:3156–3165.
          pmc: PMC111816pubmed: 10708432
        45. Ouzounis C A, Blencowe B J. Bacterial DNA replication initiation factor priA is related to proteins belonging to the ‘DEAD-box’ family.. Nucleic Acids Res 1991;19:6953.
          pmc: PMC329337pubmed: 1662369
        46. Pedersen K W, van der Meer Y, Roos N, Snijder E J. Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex.. J Virol 1999;73:2016–2026.
          pmc: PMC104444pubmed: 9971782
        47. Plagemann P G W. Lactate dehydrogenase-elevating virus and related viruses.. In: Fields B N, Knipe D M, Howley P M, editors. Fields virology. 3rd ed. Philadelphia, Pa: Lippincott-Raven Publishers; 1996. pp. 1105–1120.
        48. Poch O, Sauvaget I, Delarue M, Tordo N. Identification of four conserved motifs among the RNA dependent polymerase encoding elements.. EMBO J 1989;8:3867–3874.
          pmc: PMC402075pubmed: 2555175
        49. Salmeron J M Jr, Johnston S A. Analysis of the Kluyveromyces lactis positive regulatory gene LAC9 reveals functional homology to, but sequence divergence from, the Saccharomyces cerevisiae GAL4 gene.. Nucleic Acids Res 1986;14:7767–7781.
          pmc: PMC311795pubmed: 3022234
        50. Sambrook J, Fritsch E F, Maniatis T. Molecular cloning: a laboratory manual.. 2nd ed. Cold Spring Harbor, N.Y: Cold Spring Harbor Laboratory Press; 1989.
        51. Sawicki S G, Sawicki D L. Coronaviruses use discontinuous extension for synthesis of subgenome-length negative strands.. Adv Exp Biol Med 1995;380:499–506.
          pubmed: 8830530
        52. Snijder E J, den Boon J A, Bredenbeek P J, Horzinek M C, Rijnbrand R, Spaan W J M. The carboxyl-terminal part of the putative Berne virus polymerase is expressed by ribosomal frameshifting and contains sequence motifs which indicate that toro- and coronaviruses are evolutionarily related.. Nucleic Acids Res 1990;18:4535–4542.
          pmc: PMC331274pubmed: 2388833
        53. Snijder E J, Meulenberg J J M. The molecular biology of arteriviruses.. J Gen Virol 1998;79:961–979.
          pubmed: 9603311
        54. Snijder E J, van Tol H, Pedersen K W, Raamsman M J B, de Vries A A F. Identification of a novel structural protein of arteriviruses.. J Virol 1999;73:6335–6345.
          pmc: PMC112712pubmed: 10400725
        55. Snijder E J, Wassenaar A L M, Spaan W J M. The 5′ end of the equine arteritis virus replicase gene encodes a papainlike cysteine protease.. J Virol 1992;66:7040–7048.
          pmc: PMC240365pubmed: 1331507
        56. Snijder E J, Wassenaar A L M, Spaan W J M. Proteolytic processing of the replicase ORF1a protein of equine arteritis virus.. J Virol 1994;68:5755–5764.
          pmc: PMC236979pubmed: 8057457
        57. Snijder E J, Wassenaar A L M, van Dinten L C, Spaan W J M, Gorbalenya A E. The arterivirus nsp2 protease: an unusual cysteine protease with primary structure similarities to both papain-like and chymotrypsin-like proteases.. J Biol Chem 1995;270:16671–16676.
          pubmed: 7622476
        58. Snijder E J, Wassenaar A L M, van Dinten L C, Spaan W J M, Gorbalenya A E. The arterivirus nsp4 protease is the prototype of a novel group of chymotrypsin-like enzymes, the 3C-like serine proteases.. J Biol Chem 1996;271:4864–4871.
          pubmed: 8617757
        59. Spaan W J M, Delius H, Skinner M, Armstrong J, Rottier P J M, Smeekens S, van der Zeijst B A M, Siddell S G. Coronavirus mRNA synthesis involves fusion of non-contiguous sequences.. EMBO J 1983;2:1839–1844.
          pmc: PMC555368pubmed: 6196191
        60. Spaan W J M, Rottier P J M, Horzinek M C, van der Zeijst B A M. Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A59). Virology 1981;108:424–434.
          pmc: PMC7130792pubmed: 6258295
        61. van der Meer Y, van Tol H, Krijnse Locker J, Snijder E J. ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex.. J Virol 1998;72:6689–6698.
          pmc: PMC109868pubmed: 9658116
        62. van Dinten L C, den Boon J A, Wassenaar A L M, Spaan W J M, Snijder E J. An infectious arterivirus cDNA clone: identification of a replicase point mutation which abolishes discontinuous mRNA transcription.. Proc Natl Acad Sci USA 1997;94:991–996.
          pmc: PMC19627pubmed: 9023370
        63. van Dinten L C, Rensen S, Gorbalenya A E, Snijder E J. Proteolytic processing of the open reading frame 1b-encoded part of arterivirus replicase is mediated by nsp4 serine protease and is essential for virus replication.. J Virol 1999;73:2027–2037.
          pmc: PMC104445pubmed: 9971783
        64. van Dinten L C, Wassenaar A L M, Gorbalenya A E, Spaan W J M, Snijder E J. Processing of the equine arteritis virus replicase ORF1b protein: identification of cleavage products containing the putative viral polymerase and helicase domains.. J Virol 1996;70:6625–6633.
          pmc: PMC190703pubmed: 8794297
        65. van Marle G, Dobbe J C, Gultyaev A P, Luytjes W, Spaan W J M, Snijder E J. Arterivirus discontinuous mRNA transcription is guided by base-pairing between sense and antisense transcription-regulating sequences.. Proc Natl Acad Sci USA 1999;96:12056–12061.
          pmc: PMC18411pubmed: 10518575
        66. van Marle G, van Dinten L C, Luytjes W, Spaan W J M, Snijder E J. Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis.. J Virol 1999;73:5274–5281.
          pmc: PMC112582pubmed: 10364273
        67. Visse R, de Ruijter M, Ubbink M, Brandsma J A, van de Putte P. The first zinc-binding domain of UvrA is not essential for UvrABC-mediated DNA excision repair.. Mutat Res 1993;294:263–274.
          pubmed: 7692266
        68. Wassenaar A L M, Spaan W J M, Gorbalenya A E, Snijder E J. Alternative proteolytic processing of the arterivirus replicase ORF1a polyprotein: evidence that NSP2 acts as a cofactor for the NSP4 serine protease.. J Virol 1997;71:9313–9322.
          pmc: PMC230234pubmed: 9371590
        69. Wertman K F, Drubin D G, Botstein D. Systematic mutational analysis of the yeast ACT1 gene.. Genetics 1992;132:337–350.
          pmc: PMC1205140pubmed: 1427032
        70. Witte M M, Dickson R C. Cysteine residues in the zinc finger and amino acids adjacent to the finger are necessary for DNA binding by the LAC9 regulatory protein of Kluyveromyces lactis.. Mol Cell Biol 1988;8:3726–3733.
          pmc: PMC365429pubmed: 3146691
        71. Witte M M, Dickson R C. The C6 zinc finger and adjacent amino acids determine DNA-binding specificity and affinity in the yeast activator proteins LAC9 and PPR1.. Mol Cell Biol 1990;10:5128–5137.
          pmc: PMC361184pubmed: 2118990
        72. Wray L V Jr, Witte M M, Dickson R C, Riley M I. Characterization of a positive regulatory gene, LAC9, that controls induction of the lactose-galactose regulon of Kluyveromyces lactis: structural and functional relationship to GAL4 of Saccharomyces cerevisiae.. Mol Cell Biol 1987;7:1111–1121.
          pmc: PMC365183pubmed: 3550430
        73. Zang W Q, Veldhoen N, Romaniuk P J. Effects of zinc finger mutations on the nucleic acid binding activities of Xenopus transcription factor IIIA.. Biochemistry 1995;34:15545–15552.
          pubmed: 7492557
        Subscribe to our newsletter
        Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.