TY - JOUR
T1 - Localization and identification of sumoylated proteins in human sperm
T2 - Excessive sumoylation is a marker of defective spermatozoa
AU - Vigodner, Margarita
AU - Shrivastava, Vibha
AU - Gutstein, Leah Elisheva
AU - Schneider, Jordana
AU - Nieves, Edward
AU - Goldstein, Marc
AU - Feliciano, Miriam
AU - Callaway, Myrasol
N1 - Funding Information:
The authors thank Frank Macaluso and Juan Jimenez from the Analytical Imaging Facility (AECOM) for their cooperation and assistance with electron microscopy experiments. The authors also thank Tomer Avidor Reiss and Stephanie Blachon (Harvard Medical School) for sending slides with fly testicles. The authors also thank the Stern College for Women and Yeshiva University for supporting this research project.
Funding Information:
This study was supported in part by a grant from the Flight Attendant Medical Research Institute (to M.V.), a grant from National Institute of Child Health and Human Development, National Institute of Health R15 HD067944-01A1 (to M.V.), SIG grant for the (LTQ) Mass Spectrometer System 1S10RR019352 and the LTQ Orbitrap Velos Mass Spectrometer System 1S10RR029398.
PY - 2013/1
Y1 - 2013/1
N2 - Background Sumoylation is a type of post-translational modification that is implicated in the regulation of numerous cellular events. However, its role in the function of human sperm has not yet been characterized. Methods and Results In this study, both immunofluorescence and electron microscopy revealed that small ubiquitin-like modifiers (SUMO) SUMO1 and SUMO2/3 were highly enriched in the neck area of human sperm that is associated with the redundant nuclear envelope and were also detectable in the flagella and some head regions. Similar localization patterns of SUMO were also observed in mouse and fly sperm. Nonmotile, two-tailed, curled tailed, misshapen, microcephalic (small head) and aciphalic (no head) sperm exhibited abnormally high levels of sumoylation in their neck and tail regions relative to normal sperm. Numerous sumoylated proteins, ranging from 20 to 260 kDa, were detected via western blotting and identified by mass spectrometry, and 55 SUMO targets that were present specifically in human sperm, and not in the control fraction, corresponded to flagella proteins, proteins involved in the maturation and differentiation of sperm, heat shock proteins and important glycolytic and mitochondrial enzymes. The targets that were identified included proteins with specific functions in germ cells and sperm, such as heat shock-related 70-kDa protein 2, outer dense fiber protein 3, A-kinase anchor proteins 3 and 4, l-lactate dehydrogenase C, sperm protein associated with the nucleus on the X chromosome B/F, valosin-containing protein, seminogelins, histone H4 and ubiquitin. Coimmunoprecipitation experiments confirmed the sumoylation of semenogelin and indicated that some sperm proteins are modified by sumoylation and ubiquitination simultaneously. Conclusionsn umerous proteins are modified by sumoylation in human sperm; excessive sumoylation is a marker of defective spermatozoa.
AB - Background Sumoylation is a type of post-translational modification that is implicated in the regulation of numerous cellular events. However, its role in the function of human sperm has not yet been characterized. Methods and Results In this study, both immunofluorescence and electron microscopy revealed that small ubiquitin-like modifiers (SUMO) SUMO1 and SUMO2/3 were highly enriched in the neck area of human sperm that is associated with the redundant nuclear envelope and were also detectable in the flagella and some head regions. Similar localization patterns of SUMO were also observed in mouse and fly sperm. Nonmotile, two-tailed, curled tailed, misshapen, microcephalic (small head) and aciphalic (no head) sperm exhibited abnormally high levels of sumoylation in their neck and tail regions relative to normal sperm. Numerous sumoylated proteins, ranging from 20 to 260 kDa, were detected via western blotting and identified by mass spectrometry, and 55 SUMO targets that were present specifically in human sperm, and not in the control fraction, corresponded to flagella proteins, proteins involved in the maturation and differentiation of sperm, heat shock proteins and important glycolytic and mitochondrial enzymes. The targets that were identified included proteins with specific functions in germ cells and sperm, such as heat shock-related 70-kDa protein 2, outer dense fiber protein 3, A-kinase anchor proteins 3 and 4, l-lactate dehydrogenase C, sperm protein associated with the nucleus on the X chromosome B/F, valosin-containing protein, seminogelins, histone H4 and ubiquitin. Coimmunoprecipitation experiments confirmed the sumoylation of semenogelin and indicated that some sperm proteins are modified by sumoylation and ubiquitination simultaneously. Conclusionsn umerous proteins are modified by sumoylation in human sperm; excessive sumoylation is a marker of defective spermatozoa.
KW - defective spermatozoa
KW - redundant nuclear envelope
KW - sperm
KW - sumoylation
KW - ubiquitin
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U2 - 10.1093/humrep/des317
DO - 10.1093/humrep/des317
M3 - Article
C2 - 23077236
AN - SCOPUS:84871191616
SN - 0268-1161
VL - 28
SP - 210
EP - 223
JO - Human Reproduction
JF - Human Reproduction
IS - 1
ER -