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PubMed2000

Androcam, a Drosophila calmodulin-related protein, is expressed specifically in the testis and decorates loop kl-3 of the Y chromosome.

The Drosophila genome encodes a protein that is 68% identical to Drosophila calmodulin (Cam). We show here that this Cam-related gene is specifically expressed in the germ-line of the testis, leading to the name Androcam (Acam). Early in spermatogenesis Acam accumulates on one of the chromatin loops of the Y chromosome, kl-3. This association with kl-3 may indicate an RNA processing-related role for Acam and/or could reflect an unusual storage/assembly function hypothesized for the Y loops. After meiosis Acam is detectable in developing sperm tail cytoplasm, where at least some of the protein is not tightly associated with tubulin. Late in spermiogenesis, some Acam staining overlaps the periphery of the investment cones, actin-containing structures hypothesized to support the motor function for cytoplasmic stripping of the tail. Acam cannot be detected in mature sperm by immunolocalization, but immunoblotting established that Acam is present in sperm stored in mated females, suggesting epitope masking during final maturation. Proteins more related to Acam than Cam are present in the testes of other Drosophila species and a mammalian species, the mouse.

Animals↗

PubMed1999

Conformational and metal-binding properties of androcam, a testis-specific, calmodulin-related protein from Drosophila.

Androcam is a testis-specific protein of Drosophila melanogaster, with 67% sequence identity to calmodulin and four potential EF-hand calcium-binding sites. Spectroscopic monitoring of the thermal unfolding of recombinant calcium-free androcam shows a biphasic process characteristic of a two-domain protein, with the apo-N-domain less stable than the apo-C-domain. The two EF hands of the C-domain of androcam bind calcium cooperatively with 40-fold higher average affinity than the corresponding calmodulin sites. Magnesium competes with calcium binding [Ka(Mg) approximately 3 x 10(3) M(-1)]. Weak calcium binding is also detected at one or more N-domain sites. Compared to apo-calmodulin, apo-androcam has a smaller conformational response to calcium and a lower alpha-helical content over a range of experimental conditions. Unlike calmodulin, a tryptic cleavage site in the N-domain of apo-androcam remains trypsin sensitive in the presence of calcium, suggesting an altered calcium-dependent conformational change in this domain. The affinity of model target peptides for androcam is 10(3)-10(5) times lower than for calmodulin, and interaction of the N-domain of androcam with these peptides is significantly reduced. Thus, androcam shows calcium-induced conformational responses typical of a calcium sensor, but its properties indicate calcium sensitivity and target interactions significantly different from those of calmodulin. From the sequence differences and the altered calcium-binding properties it is likely that androcam differs from calmodulin in the conformation of residues in the second calcium-binding loop. Molecular modeling supports the deduction that there are significant conformational differences in the N-domain of androcam compared to calmodulin, and that these could affect the surface, conferring a different specificity on androcam in target interactions related to testis-specific calcium signaling functions.

Amino Acid Sequence↗

PubMed1998

Calcium-binding proteins and development.

The known roles for calcium-binding proteins in developmental signaling pathways are reviewed. Current information on the calcium-binding characteristics of three classes of cell-surface developmental signaling proteins (EGF-domain proteins, cadherins and integrins) is presented together with an overview of the intracellular pathways downstream of these surface receptors. The developmental roles delineated to date for the universal intracellular calcium sensor, calmodulin, and its targets, and for calcium-binding regulators of the cytoskeleton are also reviewed.

Animals↗