tolloid
TLD mRNA is first detected at nuclear cycle 10-11. In the central region of the embryo, only dorsally located nuclei express tld, while at the poles, both dorsally and ventrally located nuclei are labelled. The first alternation is a reduction in the staining of the cells located within 10% egg length from each pole. With further reduction, four distinct bands of expression are observed by the end of cellularization. The dorsal-most 10-20% of the embryo's circumference becomes more heavily stained.
During late stage 8, lateral patches of cells located within emerging gnathal segments are stained. During stages 10 and 11, two parallel groups of labelled cells arise. One comprises a continuous stripe of cells located between the dorsal epidermis and the amnioserosa, and a second is compared to segmentally repeated cells near the boundary between dorsal epidermis and the neurogenic region, around the tracheal pits (Schimell, 1991).
tld mutants lack a characteristic subset of dorsally derived cuticular structures from the dorsal 40% of the blastoderm fate map. Missing are cuticular specializations of the head and the fitzköpfer in the tail. There is an expansion of the ventral neurogenic ectoderm. Embryos lacking dpp activity have a more severe phenotype (Shimell, 1992).
Doubling the dpp+ gene dosage completely suppresses weak tolloid mutants and partially suppresses the phenotypes of tolloid null mutants. The function of tolloid is to increase dpp activity. tolloid, shrew
and short gastrulation genes are required to generate a gradient of dpp activity, which directly specifies the
pattern of the dorsal 40% of the embryo (Furguson, 1992).
Mutations at five loci delete specific pattern elements in the
dorsal half of the embryo and cause partial ventralization. Mutations in the genes zerknüllt and
shrew affect cell division only in the dorsalmost cells corresponding to the amnioserosa, while the
genes tolloid, screw and decapentaplegic (dpp) affect divisions in both the prospective amnioserosa
and the dorsal epidermis. In each of these mutants dorsally placed mitotic
domains are absent and this effect is correlated with an expansion and dorsal shift in the position of
more ventral domains (Arora, 1992).
Several mutant tolloid alleles have been generated and have
examined their interaction with a graded set of dpp point alleles. Some tolloid alleles act as
dominant enhancers of dpp in a trans heterozygote, and are therefore antimorphic alleles.
DPP is most
closely related to the TGF-beta superfamily members BMP-2 and BMP-4. The common
phenotype and genetic interaction data with tolloid suggests a direct physical association between these
two proteins.
However, a tolloid deficiency shows no such genetic interaction. Antimorphic dominant enhancers of dpp are missense mutations in the protease domain. Most tolloid alleles that do not interact with dpp are missense mutations in the C-terminal EGF
and C1r/s repeats, or encode truncated proteins dispersed
throughout the length of the protein (Childs, 1994 and Finelli, 1994).
The homeobox gene tinman plays a key role in the specification of Drosophila heart
progenitors and the visceral mesoderm of the midgut, both of which arise at defined
positions within dorsal areas of the mesoderm. In addition to the
heart and midgut visceral mesoderm, tinman is also required for the specification of all
dorsal body wall muscles. Thus it appears that the precursors of the heart, visceral
musculature, and dorsal somatic muscles are all specified within the same broad domain
of dorsal mesodermal tinman expression. Because of the crucial role of dpp in inducing dorsal mesodermal tinman expression and the specification of dorsal mesodermal tissues, it is of interest to determine whether other components known to function in dpp-mediated signaling events during blastoderm are also required for mesoderm induction. Screw, a second BMP2/4-related gene
product, Tolloid, a BMP1-related protein, and the zinc finger-containing protein
Schnurri, are all shown to be required to allow full levels of tinman induction during this process. screw, which encodes a second BMP2/4-related molecule, has been proposed to act synergistically with dpp to specify dorsal ectoderm and amnioserosa. Similarly, it has been shown that tolloid, which encodes a BMP1-related metalloproteinase, acts to enhance the activity of the dpp gene product during mesoderm induction. Both scw and tolloid are shown to be required for normal induction of tinman expression in the dorsal mesoderm, and in the absence of either gene activity, tinman expression in the dorsal mesoderm is reduced and segmentally interrupted. Thus scw and tld are necessary for achieving full levels of tinman induction, whereas dpp is obligatory for this event. In addition, unlike dpp mutants, mutants for scw or tld form some residual visceral mesoderm. However, heart formation is more sensitive to the activities of scw and tld and is disrupted to a similar extent as in dpp mutants. schnurri is also necessary for tinman induction in the dorsal mesoderm. The dorsal tinman domain is clearly reduced, as compared to wild-type embryos, although the levels of Tinman mRNA are close to normal. Therefore, shn may be required to enhance dpp signaling during tin induction, but significant levels of tin activation can still occur in the absence of its activity (Yin, 1998).
The BMP pathway patterns the dorsal region of the
Drosophila embryo. Using an antibody recognizing
phosphorylated Mad (pMad), signaling was followed
directly. In wild-type embryos, a biphasic activation pattern
is observed. At the cellular blastoderm stage, high pMad
levels are detected only in the dorsal-most cell rows that
give rise to amnioserosa. This accumulation of pMad
requires the ligand Screw (Scw), the Short gastrulation
(Sog) protein, and cleavage of their complex by Tolloid
(Tld). When the inhibitory activity of Sog is removed, Mad
phosphorylation is expanded. In spite of the uniform
expression of Scw, pMad expansion is restricted to the
dorsal domain of the embryo where Dpp is expressed.
This demonstrates that Mad phosphorylation requires
simultaneous activation by Scw and Dpp. Indeed, the early
pMad pattern is abolished when either the Scw receptor
Saxophone (Sax), the Dpp receptor Thickveins (Tkv), or
Dpp are removed. After germ band extension, a uniform
accumulation of pMad is observed in the entire dorsal
domain of the embryo, with a sharp border at the junction
with the neuroectoderm. From this stage onward,
activation by Scw is no longer required, and Dpp suffices
to induce high levels of pMad. In these subsequent phases
pMad accumulates normally in the presence of ectopic Sog,
in contrast to the early phase, indicating that Sog is only
capable of blocking activation by Scw and not by Dpp (Dorfman, 2001).
Thus two distinct phases of pMad
activation have been identified. The early phase requires
activation by both Scw and Dpp ligands, while the second
phase depends only on Dpp. Signaling is first detected in the cellular blastoderm embryo. While activation is observed within the dorsal-most 8-10 cell
rows, the sensitivity of the detection method fails to monitor
signaling in the rest of the dorsal domain. High signaling levels
are induced by Scw, and give rise to amnioserosa. Within the
domain where pMad is observed, graded
activation is detected, which may have the capacity to induce more than
one cell fate in the region (Dorfman, 2001).
The cardinal players in the generation of the early pMad
gradient are Scw, Tld and Sog. Tld has been suggested to generate
a sink for the active ligand, by cleaving the Sog/ligand complex. The similarity between the pMad pattern of scw and tld mutants suggests that Tld is primarily
involved in the release of Scw from the complex with Sog.
Absence of Scw, Tld or Sax abolished the early pMad
pattern while retaining the second phase, indicating that the
second phase relies only on Dpp signaling.
Similarly, overexpression of Sog eliminated only the early but
not the subsequent pMad patterns. This suggests that
Sog preferentially associates with Scw, in agreement with
previous biological assays of Sog activity.
Generation of graded patterning in the dorsal region does
not rely on restricted gene expression within this domain.
Rather, expression of genes confined to the neuroectoderm
may lead to graded distribution of their gene products within
the dorsal domain. The essential component for generation of
graded patterning appears to be Sog, which is produced only
in the neuroectoderm, but is capable of diffusing to the dorsal
region. Disruption of the normal distribution of Sog by uniform
misexpression, abolishes the early pMad activation profile (Dorfman, 2001).
This suggests that normally Sog may form a graded
distribution in the dorsal region, which is essential for
patterning. When the Sog/Scw complex is cleaved by Tld, Scw
is released and can bind either Sog or Sax. The data suggest
that in regions closer to the neuroectoderm, the levels of Sog
are high and titrate the free ligand. In the dorsal-most region
however, where Sog levels are low, the released Scw has a
greater probability of binding and activating the Sax receptor,
rather than being trapped again by Sog. Thus, the graded
distribution of Sog is critical for generating the reciprocal
distribution of Scw, and the ensuing activation profile (Dorfman, 2001).
Arora, K. and Nusslein-Volhard, C. (1992). Altered mitotic domains reveal fate map changes in Drosophila embryos mutant for zygotic dorsoventral
patterning genes. Development 114: 1003-24. PubMed Citation: 1618145
Ashe, H. L. and Levine, M. (1999). Local inhibition and long-range enhancement of Dpp signal transduction by Sog. Nature 398(6726): 427-31. PubMed Citation: 10201373
Blader, P., et al. (1997). Cleavage of the BMP-4 antagonist chordin by zebrafish tolloid. Science 278(5345): 1937-1940. PubMed Citation: 9395394
Carneiro, K., et al. (2006). Graded maternal Short gastrulation protein contributes to embryonic dorsal-ventral patterning by delayed induction.
Dev. Biol. 296(1): 203-18. 16781701
Childs, S. R. and O'Connor, M. B. (1994). Two domains of the tolloid protein contribute to its unusual genetic interaction with decapentaplegic.
Dev Biol 162: 209-20. PubMed Citation: 8125188
Clark, T. G., et al. (1999). The mammalian Tolloid-like 1 gene, Tll1, is necessary for
normal septation and positioning of the heart. Development 126(12): 2631-2642. 10331975
Dale, L., Evans, W. and Goodman, S. A. (2002). Xolloid-related: a novel BMP1/Tolloid-related metalloprotease is expressed during early Xenopus development. Mech. Dev. 119: 177-190. 12464431
Dorfman, R. and Shilo, B.-Z. (2001). Biphasic activation of the BMP pathway patterns the Drosophila embryonic dorsal region. Development 128: 965-972. 11222150
Ferguson, E. L. and Anderson, K. V. (1992). Localized enhancement and repression of the activity of the TGF-beta family member, decapentaplegic, is necessary for dorsal-ventral pattern formation in the Drosophila embryo. Development 114: 583-97. PubMed Citation: 1618130
Finelli, A. L., et al. (1994). Mutational analysis of the Drosophila tolloid gene, a human BMP-1 homolog. Development 120: 861-870. PubMed Citation: 7600963
Finelli, A. L. (1995). The tolkin gene is a tolloid/BMP-1 homologue that is essential for Drosophila development. Genetics 141: 271-281. PubMed Citation: 8536976
Goltsev, Y., Fuse, N., Frasch, M., Zinzen, R. P., Lanzaro, G. and Levine, M. (2007). Evolution of the dorsal-ventral patterning network in the mosquito, Anopheles gambiae.
Development 134(13): 2415-24. Medline abstract: 17522157
Goodman, S. A., et al. (1998). BMP1-related metalloproteinases promote the development of ventral mesoderm in early Xenopus embryos. Dev. Biol. 195(2): 144-157. PubMed Citation: 9520331
Hishida, R., et al. (1996). hch-1, a gene required for normal hatching and normal migration of a neuroblast in C. elegans, encodes a protein related to TOLLOID and BMP-1. EMBO J. 15: 4111-4122. PubMed Citation: 8861940
Holley, S. A., et al. (1996). The Xenopus dorsalizing factor noggin ventralizes Drosophila embryos by preventing DPP from activating its receptor. Cell 86: 607-617. PubMed Citation: 8752215
Jasuja, R., et al. (2006). bmp1 and mini fin are functionally redundant in regulating formation of the zebrafish dorsoventral axis. Mech. Dev. 123: 548-558. 16824737
Jazwinska, A., Rushlow, C. and Roth, S. (1999b). The role of brinker in mediating the graded response to Dpp in early Drosophila embryos. Development 126(15): 3323-3334. 10393112
Kessler, E., et al. (1996). Bone morphogenetic protein-1: the type I procollagen
C-proteinase. Science 271: 360-362. PubMed Citation: 8553073
Kirov, N., et al. (1994). The Drosophila dorsal morphogen represses the tolloid
gene by interacting with a silencer element. Mol Cell Biol 14: 713-22. PubMed Citation: 8264640
Larraín, J., et al. (2001). Proteolytic cleavage of Chordin as a switch for the dual activities of Twisted gastrulation in BMP signaling. Development 128: 4439-4447. 11714670
Lee, H. X., Ambrosio, A. L., Reversade, B. and De Robertis, E. M. (2006).
Embryonic dorsal-ventral signaling: secreted frizzled-related proteins as inhibitors of tolloid proteinases. Cell 124(1): 147-59. 16413488
Lee, H. X., Mendes, F. A., Plouhinec, J. L. and De Robertis, E. M. (2009). Enzymatic regulation of pattern: BMP4 binds CUB domains of Tolloids and inhibits proteinase activity. Genes Dev. 23(21): 2551-62. PubMed Citation: 19884260
Lee, S., et al. (1997). Transforming growth factor-beta regulation of bone morphogenetic protein-1/procollagen C-proteinase and related proteins in fibrogenic cells and keratinocytes. J. Biol. Chem. 272(30): 19059-19066
Lilja, T., et al. (2003). The CBP coactivator functions both upstream and downstream of Dpp/Screw signaling in the early Drosophila embryo. Dev. Biol. 262: 294-302. 14550792
Little, S. C. and Mullins, M. C. (2004). Twisted gastrulation promotes BMP signaling in zebrafish dorsal-ventral axial patterning. Development 131: 5825-5835. 15525664
Marques, G., et al. (1997). Production of a DPP activity gradient in the early Drosophila embryo through the opposing actions of the SOG and TLD proteins. Cell 91(3): 417-426
McPherson, C. E., Varley, J. E. and Maxwell, G. D. (2000). Expression and regulation of type I BMP receptors during early avian sympathetic
ganglion development. Dev. Biol. 221: 220-232.
Nguyen, T., et al. (1994). Characterization of tolloid-related-1: a BMP-1-like product that is required during larval and pupal stages of
Drosophila development. Dev Biol 166: 569-586
Nunes da Fonseca, R., van der Zee, M. and Roth, S. (2010). Evolution of extracellular Dpp modulators in insects: The roles of tolloid and twisted-gastrulation in dorsoventral patterning of the Tribolium embryo.
Dev. Biol. 345(1): 80-93. PubMed Citation: 20510683
Peluso, C. E., Umulis, D., Kim, Y. J., O'Connor, M. B. and Serpe, M. (2011). Shaping BMP Morphogen Gradients through Enzyme-Substrate Interactions. Dev. Cell 21(2): 375-83. PubMed Citation: 21839924
Piccolo, S., et al. (1997). Cleavage of Chordin by Xolloid metalloprotease suggests a role for proteolytic processing in the regulation of Spemann organizer activity. Cell 91(3): 407-16
Reddien, P. W., Bermange, A. L., Kicza, A. M. and Sánchez Alvarado, A.
(2007). BMP signaling regulates the dorsal planarian midline and is needed for asymmetric regeneration. Development 134(22): 4043-51. Medline abstract: 17942485
Scott, I. C., et al. (1999). Mammalian BMP-1/Tolloid-related metalloproteinases, including novel family member
mammalian Tolloid-like 2, have differential enzymatic activities and distributions of expression
relevant to patterning and skeletogenesis. Dev. Biol. 213(2): 283-300
Scott, I. C., et al. (2001). Homologues of Twisted gastrulation are extracellular cofactors in antagonism of BMP signaling. Nature 410: 475-478. 11260715
Serpe, M., et al. (2005). Matching catalytic activity to developmental function:
Tolloid-related processes Sog in order to help specify the posterior crossvein in the Drosophila wing. Development 132: 2645-2656. 15872004
Shimell, M.J., Ferguson, E.L., Childs, S.R., and O'Connor, M.B. (1991). The Drosophila dorsal-ventral patterning gene tolloid is related to human bone morphogenic protein 1. Cell 67: 469-481
Shimmi, O. and O'Connor, M. B. (2003). Physical properties of Tld, Sog, Tsg and Dpp protein interactions are predicted to help create a sharp boundary in Bmp signals during dorsoventral patterning of the Drosophila embryo. Development 130: 4673-4682. 12925593
Srinivasan, S., Rashka, K. E. and Bier, E. (2002). Creation of a Sog morphogen gradient in the Drosophila embryo. Dev. Cell 2(1): 91-101. 11782317
Suzuki, N., et al. (1996). Failure of ventral body wall closure in mouse
embryos lacking a procollagen C-proteinase encoded by Bmp1, a mammalian gene related to Drosophila tolloid. Development 122, 3587-3595
Takahara, K. Lyons, G. E. and Greenspan, D. S. (1994).
Bone morphogenetic protein-1 and a mammalian tolloid
homologue (mTld) are encoded by alternatively spliced
transcripts which are differentially expressed in some
tissues. J. Biol. Chem. 269: 32572-32578
Takahara, K., et al. (1995). Structural organization and genetic localization of the human bone morphogenetic protein 1/mammalian tolloid gene. Genomics 29: 9-15
Wardle, F. C., et al. (1999a). Regulation of BMP signaling by the BMP1/TLD-related
metalloprotease, SpAN. Dev. Biol. 206(1): 63-72
Wardle, F. C., Welch, J. V. and Dale, L. (1999b). Bone morphogenetic protein 1 regulates dorsal-ventral patterning in early Xenopus embryos by degrading chordin, a BMP4 antagonist. Mech. Dev. 86(1-2): 75-85
Xie, J. and Fisher, S. (2005). Twisted gastrulation enhances BMP signaling through
chordin dependent and independent mechanisms. Development 132: 383-391. 15604098
Yin, Z. and Frasch, M. (1998). Regulation and function of tinman during dorsal
mesoderm induction and heart specification in
Drosophila. Dev. Genet. 22(3): 187-200
Yu, K., et al. (2000). Processing of the Drosophila Sog protein creates a novel BMP inhibitory
activity. Development 127: 2143-2154. 10769238
Zhang, H., Levine, M. and Ashe, H. L. (2001). Brinker is a sequence-specific transcriptional repressor in the Drosophila embryo. Genes Dev. Vol. 15: 261-266. 11159907
Zhu, R., Santat, L. A., Markson, J. S., Nandagopal, N., Gregrowicz, J. and Elowitz, M. B. (2023). Reconstitution of morphogen shuttling circuits. Sci Adv 9(28): eadf9336. PubMed ID: 37436981
tolloid:
Biological Overview
| Evolutionary Homologs
| Regulation
| Developmental Biology
| Effects of Mutation
date revised: 10 December 2011
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