Skip to main content
SpringerLink
Log in

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

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Badami RC, Patil KB: Structure and occurrence of unusual fatty acids in minor seed oils. Prog Lipid Res 19: 119–153 (1981).

    Article  Google Scholar 

  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).

    Article  Google Scholar 

  3. Berger SL: Quantifying 32P-labeled and unlabeled nucleic acids. Methods Enzymol 152: 49–54 (1987).

    PubMed  Google Scholar 

  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).

    Article  PubMed  Google Scholar 

  5. Brody S, Mikolajczyk S: Neurospora mitochondria contain an acyl-carrier protein. Eur J Biochem 173: 353–359 (1988).

    PubMed  Google Scholar 

  6. Cattaneo R: How ‘hidden’ reading frames are expressed. Trends Biochem Sci 14: 165–167 (1989).

    Article  PubMed  Google Scholar 

  7. Cooper CL, Boyce SG, Lueking DR: Purification and characterization of Rhodobacter sphaeroides acyl carrier protein. Biochemistry 26: 2740–2746 (1987).

    PubMed  Google Scholar 

  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. 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).

    Article  Google Scholar 

  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).

    PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  13. Gavel Y, vonHeijne G: A conserved cleavage-site motif in chloroplast transit peptides. FEBS Lett 261: 455–458 (1990).

    Article  PubMed  Google Scholar 

  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. 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).

    PubMed  Google Scholar 

  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. 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).

    PubMed  Google Scholar 

  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. Hansen L: Three cDNA clones for barley leaf acyl carrier proteins I and III. Carlsberg Res Commun 52: 381–392 (1987).

    Google Scholar 

  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. 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).

    Article  PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  23. Jacobson A: Purification and fractionation of poly(A)+ RNA. Methods Enzymol 152: 254–261 (1987).

    PubMed  Google Scholar 

  24. Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucleic Acids Res 15: 9627–9640 (1987).

    PubMed  Google Scholar 

  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).

    Article  Google Scholar 

  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. 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).

    PubMed  Google Scholar 

  28. Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).

    PubMed  Google Scholar 

  29. Lee JJ, Costlow NA: A molecular titration assay to measure transcript prevalence levels. Methods Enzymol 152: 633–648 (1987).

    PubMed  Google Scholar 

  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).

    Article  PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  32. Maeda N, Smithies O: The evolution of multigene families: human haptoglobin genes. Annu Rev Genet 20: 81–108 (1986).

    Article  PubMed  Google Scholar 

  33. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  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. 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. 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).

    PubMed  Google Scholar 

  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. 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).

    PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  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. 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. 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).

    Article  PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  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).

    PubMed  Google Scholar 

  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. 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).

    Article  Google Scholar 

  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).

    Article  PubMed  Google Scholar 

  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).

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany and Plant Pathology, Michigan State University, 48824, East Lansing, MI, USA

    Katherine M. Schmid & John B. Ohlrogge

Authors
  1. Katherine M. Schmid
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John B. Ohlrogge
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schmid, K.M., Ohlrogge, J.B. A root acyl carrier protein-II from spinach is also expressed in leaves and seeds. Plant Mol Biol 15, 765–778 (1990). https://doi.org/10.1007/BF00016126

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00016126

Key words

Search

Navigation