Strongylocentrotus
purpuratus Genes in Development: Primary
mesenchyme-specific genes
SM30
Function
The SM30 gene participates in the process of
biomineralization and construction of the
embryonic spicules.
It codes for a 30,6 kDa spicule matrix protein (George et al., 1991).
There are at least two SM30 genes per haploid genome (Akasaka et al., 1994).
Protein
The SM30 protein is acidic and very similar
to the C-type lectin proteins.
It has a low concentration of basic residues (Lyz and Arg
make up 6.2% of the total protein and 3.9% of the acidic
region).
While the overall protein is acidic, its carboxyl
end is very basic. The carbohyl one-sixth of the peptide,
beginning at residue 225, contains approximately one-half
of the total basic residues. This region also comprises
the most hydrophilic portion of the protein.
The SM30 basic region may be involved in interacting
with acidic domains of itself and/or with other spicule
matrix proteins.
Another feature of the SM30 protein is its proline-rich
character. Proline, at 11% of the total residues, is
the most abundant amino acid. A short stretch of residues
beginning at amino acid 205 is extremely proline-rich.
This sequence lies between the acidic and the basic
portions of the protein and may thus serve as a hinge
region interconnecting the two domains.
The SM30 protein contains a putative hydrophobic signal
sequence at its amino end, indicating that the mature
peptide may be secreted.
The size of the SM30 protein after cleavage of the putative
signal sequence is approximately 30,6 kDa.
Benson et al.(1986), Vekatesan et al.(1986), Killian et al.(1996)
have shown that a group of matrix proteins exists around
this size. However, due to a presence of the potential
N-linked glycosilation site, the 30,6 kDa SM30 protein
may actually turn out to be modified so that it migrates
during electrophoresis as one of the largest spicule
matrix proteins (George et al., 1991).
SWISS_PROT: P28163
Subcellular location
Expression Pattern
The SM30 gene is expressed at the time of
spicule formation in the embryo.
The RNA blot analysis shows that the SM30
mRNA accumulates exclusively in mineralized tissues.
Within the embryo SM30 transcript accumulation is
restricted exclusively to the primary mesenchyme cells.
Few or no transcripts are detectable from
fertilization through the mesenchyme blastula stage.
The first appearance of SM30 transcripts corresponds to
the time when PMCs are migrating through the blastocoel.
There is a sharp increase in the levels of
the SM30 1.8-kb mRNA at middle to late mesenchyme
blastula stage (26 to 28 hr).
By the prism stage there are about
29,000 SM30 transcripts present per embryo, which
averages to approximately 480 transcripts per primary
mesenchyme cell.
The transcript levels remain high through the 3-day
pluteus stage (George et al., 1991).
mRNA level
Temporal accumulation
Method 1: Nothern blot analysis
Reference: George et al., 1991
Method 2: RNase protection assay
Reference: Akasaka et al., 1994
Stage
Egg
12h
24h
39h
48h
72h
96h
Level
-
-
- +
+
+
+
+
Spatial localization
Method: in situ hybridization
Reference: George et al., 1991