Cargando…

Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1

BACKGROUND: Anosmin-1, the protein implicated in the X-linked Kallmann's syndrome, plays a role in axon outgrowth and branching but also in epithelial morphogenesis. The molecular mechanism of its action is, however, widely unknown. Anosmin-1 is an extracellular protein which contains a cystein...

Descripción completa

Detalles Bibliográficos
Autores principales: Andrenacci, Davide, Grimaldi, Maria R, Panetta, Vittorio, Riano, Elena, Rugarli, Elena I, Graziani, Franco
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1629024/
https://www.ncbi.nlm.nih.gov/pubmed/17034626
http://dx.doi.org/10.1186/1471-2156-7-47
_version_ 1782130620979216384
author Andrenacci, Davide
Grimaldi, Maria R
Panetta, Vittorio
Riano, Elena
Rugarli, Elena I
Graziani, Franco
author_facet Andrenacci, Davide
Grimaldi, Maria R
Panetta, Vittorio
Riano, Elena
Rugarli, Elena I
Graziani, Franco
author_sort Andrenacci, Davide
collection PubMed
description BACKGROUND: Anosmin-1, the protein implicated in the X-linked Kallmann's syndrome, plays a role in axon outgrowth and branching but also in epithelial morphogenesis. The molecular mechanism of its action is, however, widely unknown. Anosmin-1 is an extracellular protein which contains a cysteine-rich region, a whey acidic protein (WAP) domain homologous to some serine protease inhibitors, and four fibronectin-like type III (FnIII) repeats. Drosophila melanogaster Kal-1 (DmKal-1) has the same protein structure with minor differences, the most important of which is the presence of only two FnIII repeats and a C-terminal region showing a low similarity with the third and the fourth human FnIII repeats. We present a structure-function analysis of the different DmKal-1 domains, including a predicted heparan-sulfate binding site. RESULTS: This study was performed overexpressing wild type DmKal-1 and a series of deletion and point mutation proteins in two different tissues: the cephalopharyngeal skeleton of the embryo and the wing disc. The overexpression of DmKal-1 in the cephalopharyngeal skeleton induced dosage-sensitive structural defects, and we used these phenotypes to perform a structure-function dissection of the protein domains. The reproduction of two deletions found in Kallmann's Syndrome patients determined a complete loss of function, whereas point mutations induced only minor alterations in the activity of the protein. Overexpression of the mutant proteins in the wing disc reveals that the functional relevance of the different DmKal-1 domains is dependent on the extracellular context. CONCLUSION: We suggest that the role played by the various protein domains differs in different extracellular contexts. This might explain why the same mutation analyzed in different tissues or in different cell culture lines often gives opposite phenotypes. These analyses also suggest that the FnIII repeats have a main and specific role, while the WAP domain might have only a modulator role, strictly connected to that of the fibronectins.
format Text
id pubmed-1629024
institution National Center for Biotechnology Information
language English
publishDate 2006
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-16290242006-10-31 Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1 Andrenacci, Davide Grimaldi, Maria R Panetta, Vittorio Riano, Elena Rugarli, Elena I Graziani, Franco BMC Genet Research Article BACKGROUND: Anosmin-1, the protein implicated in the X-linked Kallmann's syndrome, plays a role in axon outgrowth and branching but also in epithelial morphogenesis. The molecular mechanism of its action is, however, widely unknown. Anosmin-1 is an extracellular protein which contains a cysteine-rich region, a whey acidic protein (WAP) domain homologous to some serine protease inhibitors, and four fibronectin-like type III (FnIII) repeats. Drosophila melanogaster Kal-1 (DmKal-1) has the same protein structure with minor differences, the most important of which is the presence of only two FnIII repeats and a C-terminal region showing a low similarity with the third and the fourth human FnIII repeats. We present a structure-function analysis of the different DmKal-1 domains, including a predicted heparan-sulfate binding site. RESULTS: This study was performed overexpressing wild type DmKal-1 and a series of deletion and point mutation proteins in two different tissues: the cephalopharyngeal skeleton of the embryo and the wing disc. The overexpression of DmKal-1 in the cephalopharyngeal skeleton induced dosage-sensitive structural defects, and we used these phenotypes to perform a structure-function dissection of the protein domains. The reproduction of two deletions found in Kallmann's Syndrome patients determined a complete loss of function, whereas point mutations induced only minor alterations in the activity of the protein. Overexpression of the mutant proteins in the wing disc reveals that the functional relevance of the different DmKal-1 domains is dependent on the extracellular context. CONCLUSION: We suggest that the role played by the various protein domains differs in different extracellular contexts. This might explain why the same mutation analyzed in different tissues or in different cell culture lines often gives opposite phenotypes. These analyses also suggest that the FnIII repeats have a main and specific role, while the WAP domain might have only a modulator role, strictly connected to that of the fibronectins. BioMed Central 2006-10-11 /pmc/articles/PMC1629024/ /pubmed/17034626 http://dx.doi.org/10.1186/1471-2156-7-47 Text en Copyright © 2006 Andrenacci et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Andrenacci, Davide
Grimaldi, Maria R
Panetta, Vittorio
Riano, Elena
Rugarli, Elena I
Graziani, Franco
Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title_full Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title_fullStr Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title_full_unstemmed Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title_short Functional dissection of the Drosophila Kallmann's syndrome protein DmKal-1
title_sort functional dissection of the drosophila kallmann's syndrome protein dmkal-1
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1629024/
https://www.ncbi.nlm.nih.gov/pubmed/17034626
http://dx.doi.org/10.1186/1471-2156-7-47
work_keys_str_mv AT andrenaccidavide functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1
AT grimaldimariar functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1
AT panettavittorio functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1
AT rianoelena functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1
AT rugarlielenai functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1
AT grazianifranco functionaldissectionofthedrosophilakallmannssyndromeproteindmkal1