The Seminal Fluid Protease “Seminase” Regulates Proteolytic and Post-Mating Reproductive Processes
Proteases and protease inhibitors have been identified in the ejaculates of animal taxa ranging from invertebrates to mammals and form a major protein class among Drosophila melanogaster seminal fluid proteins (SFPs). Other than a single protease cascade in mammals that regulates seminal clot liquefaction, no proteolytic cascades (i.e. pathways with at least two proteases acting in sequence) have been identified in seminal fluids. In Drosophila, SFPs are transferred to females during mating and, together with sperm, are necessary for the many post-mating responses elicited in females. Though several SFPs are proteolytically cleaved either during or after mating, virtually nothing is known about the proteases involved in these cleavage events or the physiological consequences of proteolytic activity in the seminal fluid on the female. Here, we present evidence that a protease cascade acts in the seminal fluid of Drosophila during and after mating. Using RNAi to knock down expression of the SFP CG10586, a predicted serine protease, we show that it acts upstream of the SFP CG11864, a predicted astacin protease, to process SFPs involved in ovulation and sperm entry into storage. We also show that knockdown of CG10586 leads to lower levels of egg laying, higher rates of sexual receptivity to subsequent males, and abnormal sperm usage patterns, processes that are independent of CG11864. The long-term phenotypes of females mated to CG10586 knockdown males are similar to those of females that fail to store sex peptide, an important elicitor of long-term post-mating responses, and indicate a role for CG10586 in regulating sex peptide. These results point to an important role for proteolysis among insect SFPs and suggest that protease cascades may be a mechanism for precise temporal regulation of multiple post-mating responses in females.
Vyšlo v časopise:
The Seminal Fluid Protease “Seminase” Regulates Proteolytic and Post-Mating Reproductive Processes. PLoS Genet 8(1): e32767. doi:10.1371/journal.pgen.1002435
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002435
Souhrn
Proteases and protease inhibitors have been identified in the ejaculates of animal taxa ranging from invertebrates to mammals and form a major protein class among Drosophila melanogaster seminal fluid proteins (SFPs). Other than a single protease cascade in mammals that regulates seminal clot liquefaction, no proteolytic cascades (i.e. pathways with at least two proteases acting in sequence) have been identified in seminal fluids. In Drosophila, SFPs are transferred to females during mating and, together with sperm, are necessary for the many post-mating responses elicited in females. Though several SFPs are proteolytically cleaved either during or after mating, virtually nothing is known about the proteases involved in these cleavage events or the physiological consequences of proteolytic activity in the seminal fluid on the female. Here, we present evidence that a protease cascade acts in the seminal fluid of Drosophila during and after mating. Using RNAi to knock down expression of the SFP CG10586, a predicted serine protease, we show that it acts upstream of the SFP CG11864, a predicted astacin protease, to process SFPs involved in ovulation and sperm entry into storage. We also show that knockdown of CG10586 leads to lower levels of egg laying, higher rates of sexual receptivity to subsequent males, and abnormal sperm usage patterns, processes that are independent of CG11864. The long-term phenotypes of females mated to CG10586 knockdown males are similar to those of females that fail to store sex peptide, an important elicitor of long-term post-mating responses, and indicate a role for CG10586 in regulating sex peptide. These results point to an important role for proteolysis among insect SFPs and suggest that protease cascades may be a mechanism for precise temporal regulation of multiple post-mating responses in females.
Zdroje
1. SirotLKPoulsonRLMcKennaMCGirnaryHWolfnerMF 2008 Identity and transfer of male reproductive gland proteins of the dengue vector mosquito, Aedes aegypti: potential tools for control of female feeding and reproduction. Insect Biochem Mol Biol 38 176 189
2. FindlayGDYiXMaccossMJSwansonWJ 2008 Proteomics reveals novel Drosophila seminal fluid proteins transferred at mating. PLoS Biol 6 e178 doi:10.1371/journal.pbio.0060178
3. BaerBHeazlewoodJLTaylorNLEubelHMillarAH 2009 The seminal fluid proteome of the honeybee Apis mellifera. Proteomics 9 2085 2097
4. KelleherESWattsTDLaFlammeBAHaynesPAMarkowTA 2009 Proteomic analysis of Drosophila mojavensis male accessory glands suggests novel classes of seminal fluid proteins. Insect Biochem Mol Biol 39 366 371
5. MarshallJLHuestisDLHiromasaYWheelerSOppertC 2009 Identification, RNAi knockdown, and functional analysis of an ejaculate protein that mediates a postmating, prezygotic phenotype in a cricket. PLoS ONE 4 e7537 doi:10.1371/journal.pone.0007537
6. AvilaFWSirotLKLaFlammeBARubinsteinCDWolfnerMF 2011 Insect seminal fluid proteins: identification and function. Annu Rev Entomol 56 21 40
7. SouthASirotLKLewisSM 2011 Identification of predicted seminal fluid proteins in Tribolium castaneum. Insect Mol Biol 20 447 456
8. WojtczakMDietrichGJCiereszkoA 2005 Transferrin and antiproteases are major proteins of common carp seminal plasma. Fish Shellfish Immunol 19 387 391
9. MommensMWojtczakMCiereszkoABabiakI 2008 Seminal plasma proteins of Atlantic halibut (Hippoglossus hippoglossus L.). Fish Physiol Biochem 34 349 355
10. SmithJRStanfieldGM 2011 TRY-5 Is a Sperm-Activating Protease in Caenorhabditis elegans Seminal Fluid. PLoS Genet 7 e1002375 doi:10.1371/journal.pgen.1002375
11. LessleyBABrownKI 1978 Purification and properties of a proteinase inhibitor from chicken seminal plasma. Biol Reprod 19 223 234
12. Kot3owskaMKowalskiRGlogowskiJJankowskiJCiereszkoA 2005 Gelatinases and serine proteinase inhibitors of seminal plasma and the reproductive tract of turkey (Meleagris gallopavo). Theriogenology 63 1667 1681
13. MurerVSpetzJFHengstUAltroggeLMde AgostiniA 2001 Male fertility defects in mice lacking the serine protease inhibitor protease nexin-1. Proc Natl Acad Sci USA 98 3029 3033
14. DacheuxJ-LGattiJLDacheuxF 2003 Contribution of epididymal secretory proteins for spermatozoa maturation. Microsc Res Tech 61 7 17
15. PampalakisGSotiropoulouG 2007 Tissue kallikrein proteolytic cascade pathways in normal physiology and cancer. Biochim Biophys Acta 1776 22 31
16. DeanMDClarkNLFindlayGDKarnRCYiX 2009 Proteomics and comparative genomic investigations reveal heterogeneity in evolutionary rate of male reproductive proteins in mice (Mus domesticus). Mol Biol Evol 26 1733 1743
17. ChenY-WLeeM-SLuchtAChouF-PHuangW 2010 TMPRSS2, a serine protease expressed in the prostate on the apical surface of luminal epithelial cells and released into semen in prostasomes, is misregulated in prostate cancer cells. Am J Pathol 176 2986 2996
18. DeanMDFindlayGDHoopmannMRWuCCMaccossMJ 2011 Identification of ejaculated proteins in the house mouse (Mus domesticus) via isotopic labeling. BMC Genomics 12 306
19. KhanARJamesMN 1998 Molecular mechanisms for the conversion of zymogens to active proteolytic enzymes. Protein Sci 7 815 836
20. SuzukiKKiseHNishiokaJHayashiT 2007 The interaction among protein C inhibitor, prostate-specific antigen, and the semenogelin system. Semin Thromb Hemost 33 46 52
21. PoianiA 2006 Complexity of seminal fluid: a review. Behavioral Ecology and Sociobiology 60 289 310
22. KnowlesLLMarkowTA 2001 Sexually antagonistic coevolution of a postmating-prezygotic reproductive character in desert Drosophila. Proc Natl Acad Sci USA 98 8692 8696
23. WongATurchinMCWolfnerMFAquadroC 2008 Evidence for positive selection on Drosophila melanogaster seminal fluid protease homologs. Mol Biol Evol 25 497 506
24. ClarkNLAagaardJESwansonWJ 2006 Evolution of reproductive proteins from animals and plants. Reproduction 131 11 22
25. HaertyWJagadeeshanSKulathinalRJWongAKristipatiRR 2007 Evolution in the fast lane: rapidly evolving sex-related genes in Drosophila. Genetics 177 1321 1335
26. MuellerJLRipollDRAquadroCWolfnerMF 2004 Comparative structural modeling and inference of conserved protein classes in Drosophila seminal fluid. Proc Natl Acad Sci USA 101 13542 13547
27. Ravi RamKWolfnerM 2007 Seminal influences: Drosophila Acps and the molecular interplay between males and females during reproduction. Integrative and Comparative Biology 47 427 445
28. MuellerJLKristipatiRRMcGrawLABloch QaziMCSiggiaED 2005 Cross-species comparison of Drosophila male accessory gland protein genes. Genetics 171 131 143
29. GubbDSanz-ParraABarcenaLTroxlerLFullaondoA 2010 Protease inhibitors and proteolytic signalling cascades in insects. Biochimie 92 1749 1759
30. LungOTramUFinnertyCMEipper-MainsMAKalbJM 2002 The Drosophila melanogaster seminal fluid protein Acp62F is a protease inhibitor that is toxic upon ectopic expression. Genetics 160 211 224
31. WalkerMJRylettCMKeenJNAudsleyNSajidM 2006 Proteomic identification of Drosophila melanogaster male accessory gland proteins, including a pro-cathepsin and a soluble gamma-glutamyl transpeptidase. Proteome science 4 9
32. Ravi RamKSirotLKWolfnerMF 2006 Predicted seminal astacin-like protease is required for processing of reproductive proteins in Drosophila melanogaster. Proc Natl Acad Sci USA 103 18674 18679
33. SirotLKHardstoneMCHelinskiMEHRibeiroJMCKimuraM 2011 Towards a semen proteome of the dengue vector mosquito: protein identification and potential functions. PLoS Negl Trop Dis 5 e989 doi:10.1371/journal.pntd.0000989
34. KalbJMDiBenedettoAJWolfnerMF 1993 Probing the function of Drosophila melanogaster accessory glands by directed cell ablation. Proc Natl Acad Sci USA 90 8093 8097
35. HerndonLAWolfnerMF 1995 A Drosophila seminal fluid protein, Acp26Aa, stimulates egg laying in females for 1 day after mating. Proc Natl Acad Sci USA 92 10114 10118
36. Ravi RamKWolfnerMF 2007 Sustained post-mating response in Drosophila melanogaster requires multiple seminal fluid proteins. PLoS Genet 3 e238 doi:10.1371/journal.pgen.0030238
37. Bloch QaziMCWolfnerMF 2003 An early role for the Drosophila melanogaster male seminal protein Acp36DE in female sperm storage. J Exp Biol 206 3521 3528
38. AvilaFWWolfnerMF 2009 Acp36DE is required for uterine conformational changes in mated Drosophila females. Proc Natl Acad Sci USA 106 15796 15800
39. ChapmanTBanghamJVintiGSeifriedBLungO 2003 The sex peptide of Drosophila melanogaster: female post-mating responses analyzed by using RNA interference. Proc Natl Acad Sci USA 100 9923 9928
40. LiuHKubliE 2003 Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proc Natl Acad Sci USA 100 9929 9933
41. PengJChenSBüsserSLiuHHoneggerT 2005 Gradual release of sperm bound sex-peptide controls female postmating behavior in Drosophila. Curr Biol 15 207 213
42. RamKRWolfnerMF 2009 A network of interactions among seminal proteins underlies the long-term postmating response in Drosophila. Proc Natl Acad Sci USA 106 15384 15389
43. AvilaFWRavi RamKBloch QaziMCWolfnerMF 2010 Sex peptide is required for the efficient release of stored sperm in mated Drosophila females. Genetics 186 595 600
44. ParkMWolfnerMF 1995 Male and female cooperate in the prohormone-like processing of a Drosophila melanogaster seminal fluid protein. Dev Biol 171 694 702
45. HeifetzYLungOFrongilloEAWolfnerMF 2000 The Drosophila seminal fluid protein Acp26Aa stimulates release of oocytes by the ovary. Curr Biol 10 99 102
46. HeifetzYVandenbergLNCohnHIWolfnerMF 2005 Two cleavage products of the Drosophila accessory gland protein ovulin can independently induce ovulation. Proc Natl Acad Sci USA 102 743 748
47. BertramMJNeubaumDMWolfnerMF 1996 Localization of the Drosophila male accessory gland protein Acp36DE in the mated female suggests a role in sperm storage. Insect Biochem Mol Biol 26 971 980
48. NeubaumDMWolfnerMF 1999 Mated Drosophila melanogaster females require a seminal fluid protein, Acp36DE, to store sperm efficiently. Genetics 153 845 857
49. AdamsEMWolfnerMF 2007 Seminal proteins but not sperm induce morphological changes in the Drosophila melanogaster female reproductive tract during sperm storage. J Insect Physiol 53 319 331
50. BondJSBeynonRJ 1995 The astacin family of metalloendopeptidases. Protein Sci 4 1247 1261
51. ChintapalliVRWangJDowJAT 2007 Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39 715 720
52. BajajSPRapaportSIBrownSF 1981 Isolation and characterization of human factor VII. Activation of factor VII by factor Xa. J Biol Chem 256 253 259
53. GilbertD 1981 Ejaculate esterase-6 and initial sperm use by female Drosophila melanogaster. J Insect Physiol 27 641 650
54. YiallourosIKappelhoffRSchillingOWegmannFHelmsMW 2002 Activation mechanism of pro-astacin: role of the pro-peptide, tryptic and autoproteolytic cleavage and importance of precise amino-terminal processing. J Mol Biol 324 237 246
55. AmourABirdMChaudryLDeadmanJHayesD 2004 General considerations for proteolytic cascades. Biochem Soc Trans 32 15 16
56. GuevaraTYiallourosIKappelhoffRBissdorfSStöckerW 2010 Proenzyme structure and activation of astacin metallopeptidase. J Biol Chem 285 13958 13965
57. LungOWolfnerMF 2001 Identification and characterization of the major Drosophila melanogaster mating plug protein. Insect Biochem Mol Biol 31 543 551
58. Bloch QaziMCWolfnerMF 2006 Emergence of sperm from female storage sites has egg-influenced and egg-independent phases in Drosophila melanogaster. Biol Lett 2 128 130
59. SirotLKLaFlammeBASitnikJLRubinsteinCDAvilaFW 2009 Molecular social interactions: Drosophila melanogaster seminal fluid proteins as a case study. Adv Genet 68 23 56
60. AguadéM 1998 Different forces drive the evolution of the Acp26Aa and Acp26Ab accessory gland genes in the Drosophila melanogaster species complex. Genetics 150 1079 1089
61. WongATurchinMWolfnerMFAquadroCF 2011 Temporally variable selection on proteolysis related reproductive tract proteins in Drosophila. Mol Biol Evol [Published online ahead of print Sept. 22, 2011]
62. EmamiNDiamandisEP 2010 Potential role of multiple members of the kallikrein-related peptidase family of serine proteases in activating latent TGF beta 1 in semen. Biol Chem 391 85 95
63. MattssonJMValmuLLaakkonenPStenmanU-HKoistinenH 2008 Structural characterization and anti-angiogenic properties of prostate-specific antigen isoforms in seminal fluid. Prostate 68 945 954
64. Ravi RamKJiSWolfnerMF 2005 Fates and targets of male accessory gland proteins in mated female Drosophila melanogaster. Insect Biochem Mol Biol 35 1059 1071
65. CuiJSacktonKHornerVLKumarKEWolfnerMF 2008 Wispy, the Drosophila homolog of GLD-2, is required during oogenesis and egg activation. Genetics 178 2017 2029
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2012 Číslo 1
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Microenvironmental Regulation by Fibrillin-1
- Poly(ADP-Ribose) Polymerase 1 (PARP-1) Regulates Ribosomal Biogenesis in Nucleoli
- Parallel Mapping and Simultaneous Sequencing Reveals Deletions in and Associated with Discrete Inherited Disorders in a Domestic Dog Breed
- Two-Component Elements Mediate Interactions between Cytokinin and Salicylic Acid in Plant Immunity