HAP2(GCS1)-Dependent Gamete Fusion Requires a Positively Charged Carboxy-Terminal Domain

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HAP2(GCS1) is a deeply conserved sperm protein that is essential for gamete fusion. Here we use complementation assays to define major functional regions of the Arabidopsis thaliana ortholog using HAP2(GCS1) variants with modifications to regions amino(N) and carboxy(C) to its single transmembrane domain. These quantitative in vivo complementation studies show that the N-terminal region tolerates exchange with a closely related sequence, but not with a more distantly related plant sequence. In contrast, a distantly related C-terminus is functional in Arabidopsis, indicating that the primary sequence of the C-terminus is not critical. However, mutations that neutralized the charge of the C-terminus impair HAP2(GCS1)-dependent gamete fusion. Our results provide data identifying the essential functional features of this highly conserved sperm fusion protein. They suggest that the N-terminus functions by interacting with female gamete-expressed proteins and that the positively charged C-terminus may function through electrostatic interactions with the sperm plasma membrane.

Vyšlo v časopise: HAP2(GCS1)-Dependent Gamete Fusion Requires a Positively Charged Carboxy-Terminal Domain. PLoS Genet 6(3): e32767. doi:10.1371/journal.pgen.1000882
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1000882

Souhrn

Zdroje

1. SwansonWJ

VacquierVD

2002 The rapid evolution of reproductive proteins. Nat Rev Genet 3 137 144

2. JohnsonMA

von BesserK

ZhouQ

SmithE

AuxG

2004 Arabidopsis hapless mutations define essential gametophytic functions. Genetics 168 971 982

3. MoriT

KuroiwaH

HigashiyamaT

KuroiwaT

2006 GENERATIVE CELL SPECIFIC 1 is essential for angiosperm fertilization. Nat Cell Biol 8 64 71

4. von BesserK

FrankAC

JohnsonMA

PreussD

2006 Arabidopsis HAP2(GCS1) is a sperm-specific gene required for pollen tube guidance and fertilization. Development 133 4761 4769

5. SwansonR

EdlundAF

PreussD

2004 Species specificity in pollen-pistil interactions. Annu Rev Genet 38 793 818

6. BergerF

HamamuraY

IngouffM

HigashiyamaT

2008 Double fertilization - caught in the act. Trends Plant Sci 13 437 443

7. LiuY

TewariR

NingJ

BlagboroughAM

GarbomS

2008 The conserved plant sterility gene HAP2 functions after attachment of fusogenic membranes in Chlamydomonas and Plasmodium gametes. Genes Dev 22 1051 1068

8. SteeleRE

DanaCE

2009 Evolutionary history of the HAP2/GCS1 gene and sexual reproduction in metazoans. PLoS ONE 4 e7680 doi:10.1371/journal.pone.0007680

9. HiraiM

AraiM

MoriT

MiyagishimaSY

KawaiS

2008 Male fertility of malaria parasites is determined by GCS1, a plant-type reproduction factor. Curr Biol 18 607 613

10. PreussD

RheeSY

DavisRW

1994 Tetrad analysis possible in Arabidopsis with mutation of the QUARTET (QRT) genes. Science 264 1458 1460

11. SouthernJA

YoungDF

HeaneyF

BaumgartnerWK

RandallRE

1991 Identification of an epitope on the P and V proteins of simian virus 5 that distinguishes between two isolates with different biological characteristics. J Gen Virol 72 (Pt7) 1551 1557

12. AdamsSR

CampbellRE

GrossLA

MartinBR

WalkupGK

2002 New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications. J Am Chem Soc 124 6063 6076

13. WolfeKH

GouyM

YangYW

SharpPM

LiWH

1989 Date of the monocot-dicot divergence estimated from chloroplast DNA sequence data. Proc Natl Acad Sci U S A 86 6201 6205

14. BaileyCD

KochMA

MayerM

MummenhoffK

O'KaneSLJr

2006 Toward a global phylogeny of the Brassicaceae. Mol Biol Evol 23 2142 2160

15. BlagboroughAM

SindenRE

2009 Plasmodium berghei HAP2 induces strong malaria transmission-blocking immunity in vivo and in vitro. Vaccine 20 5187 5194

16. HaygoodR

2004 Sexual conflict and protein polymorphism. Evolution Int J Org Evolution 58 1414 1423

17. OkudaS

TsutsuiH

ShiinaK

SprunckS

TakeuchiH

2009 Defensin-like polypeptide LUREs are pollen tube attractants secreted from synergid cells. Nature 458 357 361

18. ShmulevitzM

DuncanR

2000 A new class of fusion-associated small transmembrane (FAST) proteins encoded by the non-enveloped fusogenic reoviruses. EMBO J 19 902 912

19. WongJL

JohnsonMA

2010 Is HAP2-GCS1 an ancestral gamete fusogen? Trends Cell Biol Jan 15. [Epub ahead of print]

20. CorcoranJA

SalsmanJ

de AntuenoR

TouhamiA

JerichoMH

2006 The p14 fusion-associated small transmembrane (FAST) protein effects membrane fusion from a subset of membrane microdomains. J Biol Chem 281 31778 31789

21. BasanezG

2002 Membrane fusion: the process and its energy suppliers. Cell Mol Life Sci 59 1478 1490

22. ChernomordikLV

KozlovMM

2008 Mechanics of membrane fusion. Nat Struct Mol Biol 15 675 683

23. WicknerW

SchekmanR

2008 Membrane fusion. Nat Struct Mol Biol 15 658 664

24. NagaharaH

Vocero-AkbaniAM

SnyderEL

HoA

LathamDG

1998 Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. Nat Med 4 1449 1452

25. HajdukiewiczP

SvabZ

MaligaP

1994 The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25 989 994

26. KonczC

SchellJ

1986 The Promoter of Tl-DNA Gene 5 Controls the Tissue-Specific Expression of Chimeric Genes Carried by a Novel Type of Agrobacterium Binary Vector. Molecular & General Genetics 204 383 396

27. CloughSJ

BentAF

1998 Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16 735 743

28. IngouffM

HamamuraY

GourguesM

HigashiyamaT

BergerF

2007 Distinct dynamics of HISTONE3 variants between the two fertilization products in plants. Curr Biol 17 1032 1037

29. OkadaT

EndoM

SinghMB