Skip to main content
Log in

Synthesis of an unusual α-zein protein is correlated with the phenotypic effects of the floury2 mutation in maize

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

The soft, starchy endosperm of the maize (Zea mays L)floury2 mutant is associated with a reduction in zein mRNA and protein synthesis, unique protein body morphology, and enhanced levels of a 70 kDa protein, that has been shown to be the maize homolog of a chaperonin found in the endoplasmic reticulum. We found an unusual α-zein protein of 24 kDa to be consistently associated with the zein fraction from floury2 mutants. Three additional α-zein proteins with molecular weights ranging from ca. 25 to 27 kDa are detected in the storage protein fraction of a high percentage of floury2 kernels and a low percentage of normal kernels in a genetically segregating population. The four proteins can be distinguished from one another by immunostaining on Western blots. Synthesis of the 24 kDa protein is regulated by Opaque2, since the 24 kDa protein is lacking in the storage protein fraction of opaque2/floury2 double mutants. The synthesis of an abnormal a-zein protein in floury2 could explain many features of the mutant, such as the abnormal protein body morphology, induction of the 70 kDa chaperonin, and hypostasis to opaque2 (o2). Although we cannot prove that the accumulation of this protein is responsible for the floury2 phenotype, we were able to detect a restriction fragment length polymorphism (RFLP) linked to the floury2 locus with a 22 kDa α-zein probe. We hypothesize that the unique characteristics of the floury2 mutant could be a response to the accumulation of a defective a-zein protein which impairs secretory protein synthesis.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Argos P, Pedersen K, Marks MD, Larkins BA (1982) A structural model for maize proteins. J Biol Chem 257:9984–9990

    Google Scholar 

  • Bass HW, Webster C, O'Brian GR, Roberts JKM, Boston RS (1992) A maize ribosome-inactivating protein is controlled by the transcriptional activator opaque2. Plant Cell 4:225–234

    Google Scholar 

  • Boston RS, Fontes EBP, Shank BB, Wrobel RL (1991) Increased expression of the maize immunoglobulin binding protein homolog b-70 in three zein regulatory mutants. Plant Cell 3:497–505

    Google Scholar 

  • Burr FA, Burr B (1982) Three mutations in Zea mays affecting zein accumulation: a comparison of zein polypeptides, in vitro synthesis and processing, mRNA levels, and genomic organization. J Cell Biol 94:201–206

    Google Scholar 

  • DiFonzo N, Fornasari E, Salamini F, Reggiane R, Soave C (1980) Interaction of the mutants floury2, opaque7 with opaque2 in the synthesis of endosperm proteins. J Hered 71:397–402

    Google Scholar 

  • Emerson RA, Beadle GW, Frazer AC (1935) A summary of linkage studies in maize. Cornell University Agricultural Experimental Station Report 180

  • Flynn GC, Chappell TG, Rothman JE (1989) Peptide binding and release by proteins implicated as catalysts of protein assembly. Science 245:385–390

    Google Scholar 

  • Fontes EBP, Shank BB, Moose RL, O'Brian GR, Wurtzel ET, Boston RS (1991) Characterization of an immunoglobulin binding protein homolog in the maize floury2 endosperm mutant. Plant Cell 3:483–496

    Google Scholar 

  • Galante E, Vitale A, Manzocchi L, Soave C, Salamini F (1983) Genetic control of a membrane component and zein deposition in maize endosperm. Mol Gen Genet 192:316–321

    Google Scholar 

  • Geetha KB, Lending CR, Lopes MA, Wallace JC, Larkins BA (1991) opaque2 modifiers increase γ-zein synthesis and alter its spatial distribution in maize endosperm. Plant Cell 3:1207–1219

    Google Scholar 

  • Geraghty DE, Peifer MA, Rubenstein I, Messing J (1981) The primary structure of a plant storage protein. Nucleic Acids Res 9:5163–5174

    Google Scholar 

  • Hard FU, Martin J (1992) Protein folding in the cell: the role of molecular chaperones Hsp70 and Hsp60. Annu Rev Biophys Biomol Struct 21:293–322

    Google Scholar 

  • Hastings H, Bonanomi S, Soave C, Di Fonzo N, Salamini F (1984) Mapping genes for minor zein SDS subunits and revision of zein genes nomenclature. Genet Agric 38:447–464

    Google Scholar 

  • Jones RA (1978) Effects of floury2 locus on zein accumulation and RNA metabolism during maize endosperm development. Biochem Genet 16:27–38

    Google Scholar 

  • Knecht DA, Dimond RL (1984) Visualization of antigenic proteins on western blots. Anal Biochem 136:180–184

    Google Scholar 

  • Kodrzycki R, Boston R, Larkins BA (1989) The opaque2 mutation of maize differentially reduces zein gene transcription. The Plant Cell 1:105–114

    Google Scholar 

  • Kreike CM, Koning JRA, Krens FA (1990) Non-radioactive detection of single-copy DNA-DNA hybrids. Plant Mol Biol Rep 8:172–179

    Google Scholar 

  • Kreis M, Shewry PR (1989) Unusual features of cereal seed protein structure and evolution. BioEssays 10:201–207

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Lee KH, Jones RA, Dalby A, Tsai C (1976) Genetic regulation of storage protein content in maize endosperm. Biochem Genet 14:641–650

    Google Scholar 

  • Lending CR, Larkins BA (1989) Changes in the zein composition of protein bodies during maize endosperm development. Plant Cell 1:1011–1023

    Google Scholar 

  • Lending CR, Larkins BA (1993) Effect of the floury2 locus on protein body formation during maize endosperm development. Protoplasma 171:123–133

    Google Scholar 

  • Lending CR, Kriz AL, Larkins BA, Bracker CE (1988) Structure of maize protein bodies and immunocytochemical localization of zeins. Protoplasma 143:51–62

    Google Scholar 

  • Lohmer S, Maddaloni M, Motto M, Di Fonzo N, Hartings H, Salamini F, Thompson RD (1991) The maize regulatory locus Opaque2, encodes a DNA-binding protein which activates transcription of the b-32 gene. EMBO J 10:617–624

    Google Scholar 

  • Lopes MA, Larkins BA (1991) γ-zein content is related to endosperm modification in quality protein maize (QPM). Crop Sci 31:1655–1662

    Google Scholar 

  • Marks MD, Larkins BA (1982) Analysis of sequence micro-heterogeneity among zein mRNAs. J Biol Chem 257:9976–9983

    Google Scholar 

  • Marocco A, Santucci A, Cerioli S, Motto M, Di Fonzo N, Thompson R, Salamini F (1991) Three high-lysine mutations control the level of ATP-binding HSP70-like proteins in the maize endosperm. Plant Cell 3:507–515

    Google Scholar 

  • McCreery T, Helentjaris T (1993a) Production of hybridization probes by the PCR utilizing digoxigenin-modified nucleotides. Methods Mol Biol 28:67–71

    Google Scholar 

  • McCreery T, Helentjaris T (1993b) Hybridization of digoxigeninlabelled probes to Southern blots and detection by chemiluminescence. Methods Mol Biol 28:107–112

    Google Scholar 

  • McDonald RJ, Swift GH, Przybyla AE, Chirgwin JM (1987) Isolation of RNA using guanidinium salts. Methods Enzymol 152:219–227

    Google Scholar 

  • Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832

    Google Scholar 

  • Motto M, Di Fonzo N, Hartings H, Maddaloni M, Salamini F, Soave C, Thompson RD (1989) Regulatory genes affecting maize storage proteins. Oxford Surveys Plant Mol Cell Biol 6:87–114

    Google Scholar 

  • Pedersen K, Devereux DR, Wilson E, Sheldon E, Larkins BA (1982) Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell 29:1015–1026

    Google Scholar 

  • Saghai-Maroof MA, Soliman KM, Jorgensen R, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018

    Google Scholar 

  • Schmidt RJ (1993) Opaque2, and zein gene expression. In: Verma DPS (ed) Control of plant gene expression. CRC Press, Boca Raton, Florida, pp 337–355

    Google Scholar 

  • Soave C, Salamini F (1984) Organization and regulation of zein genes in maize endosperm. Phil Trans R Soc Lond B 304:341–347

    Google Scholar 

  • Soave C, Dossena S, Lorenzoni C, Di Fonzo N, Salamini F (1978) Expressivity of the floury2 allele at the level of zein molecular components. Maydica 23:145–152

    Google Scholar 

  • Spena A, Viotti A, Pirrota V (1982) A homologous repetitive block structure underlies the heterogeneity of heavy and light chain zein genes. EMBO J 1:1589–1594

    Google Scholar 

  • Todorov IN, Shen RA, Zhelyabovskaya SM, Galkin AP (1977) Control of RNA synthesis in rat liver: some features of RNA synthesis during prolonged inhibition of protein synthesis. Biochemistry 41:1509–1519

    Google Scholar 

  • Todorov IN, Smalko Y, Galkin AP (1978) State of the polyribosomes as a reflection of the functional interaction of the systems of translation and transcription during the restoration of protein biosynthesis inhibited by cycloheximide. Biochemistry 42:1693–1700

    Google Scholar 

  • Ueda T, Waverczak W, Ward K, Sher N, Ketudat M, Schmidt R, Messing J (1992) Mutations of the 22- and 27-kDa zein promoters affect transactivation by the Opaque2, protein. Plant Cell 4:701–709

    Google Scholar 

  • Wallace JC, Lopes MA, Paiva E, Larkins BA (1990) New methods for extraction and quantitation of zeins reveal a high content of 7-zein in modified opaque2 maize. Plant Physiol 92:191–196

    Google Scholar 

  • Zhang F, Boston RS (1993) Increases in binding protein (BiP) accompany changes in protein body morphology in three high-lysine mutants of maize. Protoplasma 171:142–152

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by R. G. Herrmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lopes, M.A., Coleman, C.E., Kodrzycki, R. et al. Synthesis of an unusual α-zein protein is correlated with the phenotypic effects of the floury2 mutation in maize. Molec. Gen. Genet. 245, 537–547 (1994). https://doi.org/10.1007/BF00282216

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00282216

Key words

Navigation