Cloning and expression of a cDNA that comprises part of the gene coding for a spore coat protein of Dictyostelium discoideum

1984 ◽  
Vol 4 (11) ◽  
pp. 2273-2278
Author(s):  
B C Dowds ◽  
W F Loomis
Keyword(s):  
Dictyostelium Discoideum ◽  
Cdna Clone ◽  
Single Gene ◽  
Polyclonal Antiserum ◽  
Cloning And Expression ◽  
Spore Coat ◽  
Coat Proteins ◽  
Developmentally Regulated ◽  
Gene Coding ◽  
Spore Coat Proteins

The three major spore coat proteins of Dictyostelium discoideum are developmentally regulated, cell-type-specific proteins. They are packaged in prespore vesicles and then secreted to form the outer layer of spore coats. We have isolated a cDNA clone from the gene coding for one of these proteins, SP96, a glycoprotein of 96,000 daltons. We screened the cDNA bank by the method of hybrid select translation followed by immunoprecipitation of the translation products with SP96-specific polyclonal antiserum. We found that the gene was first transcribed into stable mRNA a few hours before the time of detection of SP96 synthesis and that the mRNA, like the protein, accumulated specifically in prespore cells and spores. SP96 constituted the same proportion of newly synthesized protein as the proportion of its message in polyadenylated RNA. SP96 appeared to be encoded by a single gene as judged by Southern blot analysis of digested genomic DNA hybridized to the cDNA clone.

scholarly journals Cloning and expression of a cDNA that comprises part of the gene coding for a spore coat protein of Dictyostelium discoideum.

1984 ◽  
Vol 4 (11) ◽  
pp. 2273-2278 ◽  
Author(s):  
B C Dowds ◽  
W F Loomis
Keyword(s):  
Dictyostelium Discoideum ◽  
Cdna Clone ◽  
Single Gene ◽  
Polyclonal Antiserum ◽  
Cloning And Expression ◽  
Spore Coat ◽  
Coat Proteins ◽  
Developmentally Regulated ◽  
Gene Coding ◽  
Spore Coat Proteins

The three major spore coat proteins of Dictyostelium discoideum are developmentally regulated, cell-type-specific proteins. They are packaged in prespore vesicles and then secreted to form the outer layer of spore coats. We have isolated a cDNA clone from the gene coding for one of these proteins, SP96, a glycoprotein of 96,000 daltons. We screened the cDNA bank by the method of hybrid select translation followed by immunoprecipitation of the translation products with SP96-specific polyclonal antiserum. We found that the gene was first transcribed into stable mRNA a few hours before the time of detection of SP96 synthesis and that the mRNA, like the protein, accumulated specifically in prespore cells and spores. SP96 constituted the same proportion of newly synthesized protein as the proportion of its message in polyadenylated RNA. SP96 appeared to be encoded by a single gene as judged by Southern blot analysis of digested genomic DNA hybridized to the cDNA clone.

The PsB glycoprotein complex is secreted as a preassembled precursor of the spore coat in Dictyostelium discoideum

1994 ◽  
Vol 107 (9) ◽  
pp. 2567-2579 ◽  
Author(s):  
N. Watson ◽  
V. McGuire ◽  
S. Alexander
Keyword(s):  
Extracellular Matrix ◽  
Dictyostelium Discoideum ◽  
Cellular Localization ◽  
Direct Result ◽  
Spore Coat ◽  
Coat Proteins ◽  
Multiprotein Complex ◽  
Post Translational Modification ◽  
Developmentally Regulated ◽  
Spore Coat Proteins

The PsB glycoprotein in Dictyostelium discoideum is one of a diverse group of developmentally regulated, prespore-cell-specific proteins, that contain a common O-linked oligosaccharide. This post-translational modification is dependent on the wild-type modB allele. The PsB protein exists as part of a multiprotein complex of six different proteins, which have different post-translational modifications and are held together by both covalent and non-covalent interactions (Watson et al. (1993). J. Biol. Chem. 268, 22634–22641). In this study we have used microscopic and biochemical analyses to examine the cellular localization and function of the PsB complex during development. We found that the PsB complex first accumulates in prespore vesicles in slug cells and is secreted later during culmination and becomes localized to both the extracellular matrix of the apical spore mass of mature fruiting bodies and to the inner layer of the spore coat. The PsB associated with the spore coat is covalently bound by disulfide bridges. The PsB protein always exists in a multiprotein complex, but the composition of the PsB complex changes during secretion and spore maturation. Some of the PsB complex proteins have been identified as spore coat proteins. These data demonstrate that some of the proteins that form the spore coat exist as a preassembled precursor complex. The PsB complex is secreted in a developmentally regulated manner during the process of spore differentiation, at which time proteins of the complex, as well as additional spore coat proteins, become covalently associated in at least two forms of extracellular matrix: the interspore matrix and the spore coat. These and other studies show that proteins with modB dependent O-linked oligosaccharides are involved in a wide variety of processes underlying morphogenesis in this organism. These developmental processes are the direct result of cellular mechanisms regulating protein targeting, assembly and secretion, and the assembly of specific extracellular matrices.

Phosphorylation of spore coat proteins of Dictyostelium discoideum

1983 ◽  
Vol 61 (9) ◽  
pp. 996-1001 ◽  
Author(s):  
Teshome Akalehiywot ◽  
Chi-Hung Siu
Keyword(s):  
Electrophoretic Mobility ◽  
Dictyostelium Discoideum ◽  
Cellular Proteins ◽  
Developmental Cycle ◽  
Spore Coat ◽  
Coat Proteins ◽  
Phosphoamino Acid ◽  
Molecular Weights ◽  
Phosphorylated Proteins ◽  
Spore Coat Proteins

Phosphorylation of cellular proteins was studied during development of Dictyostelium discoideum. In the second half of the developmental cycle, two heavily phosphorylated proteins were observed together with a limited number of minor phosphorylated proteins. The electrophoretic mobility of these two phosphoproteins corresponded to two of the major spore coat glycoproteins, with apparent molecular weights of 103 000 and 80 000. They were found to be externalized and associated with the spore coat during spore formation. Phosphoserine was the predominant phosphoamino acid in both cases. These two phosphoproteins thus serve as excellent markers for the differentiation of prespore cells in D. discoideum.

Structural Roles of the Spore Coat Proteins in Dictyostelium discoideum

1994 ◽  
Vol 166 (2) ◽  
pp. 823-825 ◽  
Author(s):  
Kathy Fosnaugh ◽  
Danny Fuller ◽  
William F. Loomis
Keyword(s):  
Dictyostelium Discoideum ◽  
Spore Coat ◽  
Coat Proteins ◽  
Spore Coat Proteins

scholarly journals Synthesis of spore proteins during development of Dictyostelium discoideum

1983 ◽  
Vol 216 (3) ◽  
pp. 567-574 ◽  
Author(s):  
D G Wilkinson ◽  
J Wilson ◽  
B D Hames
Keyword(s):  
Dictyostelium Discoideum ◽  
Spore Coat ◽  
Coat Proteins ◽  
Peak Period ◽  
Temporal Gene Expression ◽  
Stage Of Development ◽  
Time Period ◽  
Mexican Hat ◽  
Spore Proteins ◽  
Spore Coat Proteins

The pattern of synthesis of the spore coat proteins during development of Dictyostelium discoideum has been determined by using immunoprecipitation with spore protein antibody. SP170, SP103, ‘SP94’, SP82, SP76 and SP55 are all first synthesized just prior to the ‘Mexican hat’ stage of development (16-18h), but the synthesis of SP72 is delayed. This protein is apparently synthesized as a precursor, P66, which is modified during spore maturation to yield SP72. The nature of the modification is unknown. At their peak period of synthesis during early culmination (18-20h), the spore coat proteins account for 5-9% of total protein synthesis. Shortly after synthesis, these proteins are inserted into the spore coat, where all except SP103 become disulphide-cross-linked during the period 24-30h. SP3 does not accumulate until disulphide-cross-linking of the major spore coat proteins occurs and is itself disulphide-cross-linked into the spore coat. Several additional proteins that are accumulated during development have also been identified, namely P31, P25, P21 and P18. P25 first appears at 18-20h and then continues to be made throughout development. P31 synthesis begins at 12-14h and then largely ceases after approx. 20 h of development. The genes for both P21 and P18 are first expressed early in development, starting at 9-12h. P21 synthesis ceases at approx. 14h, but P18 continues to be synthesized throughout the rest of development. The marked differences in the time period of accumulation of these proteins compared with the co-ordinated syntheses of SP170, SP103, ‘SP94’, SP82, SP76 and SP55 provide a useful system for analysis of the mechanism of temporal gene expression during development.

Spore coat proteins of Dictyostelium discoideum are packaged in prespore vesicles

1983 ◽  
Vol 99 (2) ◽  
pp. 437-446 ◽  
Author(s):  
Kevin M. Devine ◽  
John E. Bergmann ◽  
William F. Loomis
Keyword(s):  
Dictyostelium Discoideum ◽  
Spore Coat ◽  
Coat Proteins ◽  
Spore Coat Proteins

scholarly journals Phosphorylation of spore coat proteins during development of Dictyostelium discoideum

1983 ◽  
Vol 212 (3) ◽  
pp. 699-703 ◽  
Author(s):  
S J Delaney ◽  
D G Wilkinson ◽  
B D Hames
Keyword(s):  
Coat Protein ◽  
Dictyostelium Discoideum ◽  
Spore Coat ◽  
Coat Proteins ◽  
Cellular Slime Mould ◽  
Slime Mould ◽  
Cellular Slime ◽  
Spore Coat Proteins ◽  
Spore Coat Protein

Immunological evidence is presented which confirms that pp95, one of the major phosphoproteins accumulated in development of the cellular slime mould Dictyostelium discoideum, is identical with spore coat protein SP13. The site of phosphorylation is identified as phosphoserine. The second major phosphorylated component, pp74, corresponds to two co-migrating spore coat proteins known collectively as SP74.

scholarly journals SP75 is encoded by the DP87 gene and belongs to a family of modular Dictyostelium discoideum outer layer spore coat proteins

Microbiology ◽  
1996 ◽  
Vol 142 (8) ◽  
pp. 2227-2243 ◽  
Author(s):  
C. M. West ◽  
J. Mao ◽  
H. van der Wel ◽  
G. W. Erdos ◽  
Y. Zhang
Keyword(s):  
Dictyostelium Discoideum ◽  
Outer Layer ◽  
Spore Coat ◽  
Coat Proteins ◽  
Spore Coat Proteins
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