Plant Molecular Biology

, Volume 15, Issue 5, pp 765–778 | Cite as

A root acyl carrier protein-II from spinach is also expressed in leaves and seeds

  • Katherine M. Schmid
  • John B. Ohlrogge


During the synthesis of fatty acids and their utilization in plastids, fatty acyl moieties are linked to acyl carrier protein (ACP). In contrast to previously cloned organ-specific ACP isoforms, we have now isolated a cDNA clone for a potentially constitutive ACP isoform from a spinach root library. Identity between the amino acid sequence encoded by this cDNA and N-terminal sequence data for ACP-II protein from spinach leaf indicates that the root cDNA encodes ACP-II. The deduced amino acid sequence for ACP-II shows 62% identity with spinach leaf ACP-I. Southern analysis suggests that multiple ACP genes or pseudogenes occur in the spinach genome. High-stringency northern blot analysis and RNase protection studies confirm that, within the region encoding the mature ACP-II, the cloned ACP sequence is expressed in leaves and seeds as well as in roots. Quantitative RNase protection data indicate that the ratio of ACP-I and ACP-II mRNA sequences in leaf is similar to the ratio of the two proteins.

Key words

acyl carrier protein differential expression fatty acid synthesis nuclear-encoded chloroplast proteins sequence homology Spinacia oleracea 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Badami RC, Patil KB: Structure and occurrence of unusual fatty acids in minor seed oils. Prog Lipid Res 19: 119–153 (1981).CrossRefGoogle Scholar
  2. 2.
    Battey JF, Ohlrogge JB: Evolutionary and tissue-specific control of expression of multiple acyl-carrier protein isoforms in plants and bacteria. Planta 180: 352–360 (1990).CrossRefGoogle Scholar
  3. 3.
    Berger SL: Quantifying 32P-labeled and unlabeled nucleic acids. Methods Enzymol 152: 49–54 (1987).PubMedGoogle Scholar
  4. 4.
    Breitbart RE, Andreadis A, Nadal-Ginard B: Alternative splicing: A ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Annu Rev Biochem 56: 467–495 (1987).CrossRefPubMedGoogle Scholar
  5. 5.
    Brody S, Mikolajczyk S: Neurospora mitochondria contain an acyl-carrier protein. Eur J Biochem 173: 353–359 (1988).PubMedGoogle Scholar
  6. 6.
    Cattaneo R: How ‘hidden’ reading frames are expressed. Trends Biochem Sci 14: 165–167 (1989).CrossRefPubMedGoogle Scholar
  7. 7.
    Cooper CL, Boyce SG, Lueking DR: Purification and characterization of Rhodobacter sphaeroides acyl carrier protein. Biochemistry 26: 2740–2746 (1987).PubMedGoogle Scholar
  8. 8.
    Covey SN, Grierson D: The measurement of plant polyadenylic acid by hydridisation with radioactive polyuridylic acid. Planta 131: 75–79 (1976).Google Scholar
  9. 9.
    Dean C, Pichersky E, Dunsmuir P: Structure, evolution, and regulation of Rbc S genes in higher plants. Ann Rev Plant Physiol Plant Mol Biol 40: 415–439 (1989).CrossRefGoogle Scholar
  10. 10.
    Dean C, Tamaki S, Dunsmuir P, Favreau M, Katayama C, Dooner H, Bedbrook J: mRNA transcripts of several plant genes are polyadenylated at multiple sites in vivo. Nucleic Acids Res 14: 2229–2240 (1986).PubMedGoogle Scholar
  11. 11.
    DeSilva J, Loader NM, Jarman C, Windust JHC, Hughes SG, Safford R: The isolation and sequence analysis of two seed-expressed acyl carrier protein genes from Brassica napus. Plant Mol Biol 14: 537–548 (1990).PubMedGoogle Scholar
  12. 12.
    Galau GA, Legocki AB, Greenway SC, DureIII LS: Cotton messenger RNA sequences exist in both polyadenylated and nonpolyadenylated forms. J Biol Chem 256: 2551–2560 (1981).PubMedGoogle Scholar
  13. 13.
    Gavel Y, vonHeijne G: A conserved cleavage-site motif in chloroplast transit peptides. FEBS Lett 261: 455–458 (1990).CrossRefPubMedGoogle Scholar
  14. 14.
    Gilman M: Ribonuclease protection assay. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Smith JA, Struhl K, Seidman JG (eds) Current Protocols in Molecular Biology, pp. 4.7.1–4.7.8. John Wiley & Sons, New York (1987).Google Scholar
  15. 15.
    Guerra DJ, Ohlrogge JB: Partial purification and characterization of two forms of malonyl-coenzyme A: acyl carrier protein transacylase from soybean leaf tissue. Arch Biochem Biophys 246: 274–285 (1986).PubMedGoogle Scholar
  16. 16.
    Guerra DJ, Ohlrogge JB, Frentzen M: Activity of acyl carrier protein isoforms in reactions of plant fatty acid metabolism. Plant Physiol 82: 448–453 (1986).Google Scholar
  17. 17.
    Hallick RB, Chelm BK, Gray PW, OrozcoJr. EM: Use of aurintricarboxylic acid as an inhibitor of nucleases during nucleic acid isolation. Nucleic Acids Res 4: 3055–3064 (1977).PubMedGoogle Scholar
  18. 18.
    Hannapel DJ, Ohlrogge JB: Regulation of acyl carrier protein messenger RNA levels during seed and leaf development. Plant Physiol 86: 1174–1178 (1988).Google Scholar
  19. 19.
    Hansen L: Three cDNA clones for barley leaf acyl carrier proteins I and III. Carlsberg Res Commun 52: 381–392 (1987).Google Scholar
  20. 20.
    Hansen L, vonWettstein-Knowles P: Acyl carrier proteins of barley seedling leaves and caryopses. In: Biacs PA, Gruiz K, Kremmer T (eds) Biological Role of Plant Lipids, pp. 367–370. Akademiai Klado, Budapest; Plenum, New York (1980).Google Scholar
  21. 21.
    Harada JJ, Barker SJ, Goldberg RB: Soybean β-conglycinin genes are clustered in several DNA regions and are regulated by transcriptional and posttranscriptional processes. Plant Cell 1: 415–425 (1989).CrossRefPubMedGoogle Scholar
  22. 22.
    Holak TA, Nilges M, Prestegard JH, Gronenborn AM, Clore GM: Three-dimensional structure of acyl carrier protein in solution determined by nuclear magnetic resonance and the combined use of dynamical simulated annealing and distance geometry. Eur J Biochem 175: 9–15 (1988).PubMedGoogle Scholar
  23. 23.
    Jacobson A: Purification and fractionation of poly(A)+ RNA. Methods Enzymol 152: 254–261 (1987).PubMedGoogle Scholar
  24. 24.
    Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucleic Acids Res 15: 9627–9640 (1987).PubMedGoogle Scholar
  25. 25.
    Keegstra K, Olsen LJ: Chloroplastic precursors and their transport across the envelope membranes. Annu Rev Plant Physiol Plant Mol Biol 40: 471–501 (1989).CrossRefGoogle Scholar
  26. 26.
    Krebbers E, Herdies L, DeClercq A, Seurinck J, Leemans J, VanDamme J, Segura M, Gheysen G, VanMontagu M, Vandekerckhove J: Determination of the processing sites of an Arabidopsis 2S albumin and characterization of the complete gene family. Plant Physiol 87: 859–866 (1988).Google Scholar
  27. 27.
    Kuo TM, Ohlrogge JB: Acylation of plant acyl carrier proteins by acyl-acyl carrier protein synthetase from Escherichia coli. Arch Biochem Biophys 230: 110–116 (1984).PubMedGoogle Scholar
  28. 28.
    Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).PubMedGoogle Scholar
  29. 29.
    Lee JJ, Costlow NA: A molecular titration assay to measure transcript prevalence levels. Methods Enzymol 152: 633–648 (1987).PubMedGoogle Scholar
  30. 30.
    Leff SE, Rosenfeld MG, Evans RM: Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem 55: 1091–1117 (1986).CrossRefPubMedGoogle Scholar
  31. 31.
    Leutwiler LS, Meyerowitz EM, Tobin EM: Structure and expression of three light-harvesting chlorophyll a/b-binding protein genes in Arabidopsis thaliana. Nucleic Acids Res 14: 4051–4064 (1986).PubMedGoogle Scholar
  32. 32.
    Maeda N, Smithies O: The evolution of multigene families: human haptoglobin genes. Annu Rev Genet 20: 81–108 (1986).CrossRefPubMedGoogle Scholar
  33. 33.
    Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).Google Scholar
  34. 34.
    Myers RM, Larin Z, Maniatis T: Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA: DNA duplexes. Sciences 230: 1242–1246 (1985).Google Scholar
  35. 35.
    Ohlrogge JB: Biochemistry of plant acyl carrier proteins. In: Stumpf PK (ed) Lipids: Structure and Function, pp. 137–157. The Biochemistry of Plants, Vol. 9. Academic Press, New York (1987).Google Scholar
  36. 36.
    Ohlrogge JB, Kuo TM: Plants have isoforms for acyl carrier protein that are expressed differently in different tissues. J Biol Chem 260: 8032–8037 (1985).PubMedGoogle Scholar
  37. 37.
    Polans NO, Weeden NF, Thompson WF: Inheritance, organization, and mapping of rbc S and cab multigene families in pea. Proc Natl Acad Sci USA 82: 5083–5087 (1985).Google Scholar
  38. 38.
    Post-Beittenmiller MA, Hlousek-Radojcic A, Ohlrogge JB: DNA sequence of a genomic clone encoding an Arabidopsis acyl carrier protein (ACP). Nucleic Acids Res 17: 1777 (1989).PubMedGoogle Scholar
  39. 39.
    Rose RE, DeJesus CE, Moylan SL, Ridge NP, Scherer DE, Knauf VC: The nucleotide sequence of a cDNA clone encoding acyl carrier protein (ACP) from Brassica campestris seeds. Nucleic Acids Res 15: 7197 (1987).PubMedGoogle Scholar
  40. 40.
    Safford R, Windust JHC, Lucas C, DeSilva J, James CM, Hellyer A, Smith CG, Slabas AR, Hughes SG: Plastid-localised seed acyl-carrier protein of Brassica napus in encoded by a distinct, nuclear multigene family. Eur J Biochem 174: 287–295 (1988).PubMedGoogle Scholar
  41. 41.
    Sato N: Nucleotide sequence and expression of the phytochrome tochrome gene in Pisum sativum: Differential regulation by light of multiple transcripts. Plant Mol Biol 11: 697–710 (1988).Google Scholar
  42. 42.
    Scherer DE, Knauf VC: Isolation of a cDNA clone for the acyl carrier protein-I of spinach. Plant Mol Biol 9: 127–134 (1987).Google Scholar
  43. 43.
    Sugita M, Manzara T, Pichersky E, Cashmore A, Gruissem W: Genomic organization, sequence analysis and expression of all five genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from tomato. Mol Gen Genet 209: 247–256 (1987).CrossRefPubMedGoogle Scholar
  44. 44.
    Tonnet ML, Green AG: Characterization of the seed and leaf lipids of high and low linolenic acid flax geno-types. Arch Biochem Biophys 252: 646–654 (1987).PubMedGoogle Scholar
  45. 45.
    Vanaman TC, Wakil SJ, Hill RL: The complete amino acid sequence of the acyl carrier protein of Escherichia coli. J Biol Chem 243: 6420–6431 (1968).PubMedGoogle Scholar
  46. 46.
    Vrati S, Mann DA, Reed KC: Alkaline northern blots: Transfer of RNA from agarose gels to Zeta-Probe membrane in dilute NaOH. Mol Biol Rep 1: 1–4 (1987).Google Scholar
  47. 47.
    Wang X-M, Norman HA, St.John JB, Yin T, Hildebrand DF: Comparison of fatty acid composition in tissues of low linolenate mutants of soybean. Phytochemistry 28: 411–414 (1989).CrossRefGoogle Scholar
  48. 48.
    Werneke JM, Chatfield JM, Ogren WL: Alternative mRNA splicing generates the two ribulose-bisphosphate carboxylase/oxygenase activase polypeptides in spinach and Arabidopsis. Plant Cell 1: 815–825 (1989).CrossRefPubMedGoogle Scholar
  49. 49.
    Wilkins TA, Raikhel NV: Expression of rice lectin in governed by two temporally and spatially regulated mRNAs in developing embryos. Plant Cell 1: 541–549 (1989).CrossRefPubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1990

Authors and Affiliations

  • Katherine M. Schmid
    • 1
  • John B. Ohlrogge
    • 1
  1. 1.Department of Botany and Plant PathologyMichigan State UniversityEast LansingUSA

Personalised recommendations