title: A MEIG1/PACRG complex in the manchette is essential for building the sperm flagella.
abstract: A key event in the process of spermiogenesis is the formation of the flagella, which enables sperm to reach eggs for fertilization. Yeast two-hybrid studies revealed that meiosis-expressed gene 1 (MEIG1) and Parkin co-regulated gene (PACRG) interact, and that sperm-associated antigen 16, which encodes an axoneme central apparatus protein, is also a binding partner of MEIG1. In spermatocytes of wild-type mice, MEIG1 is expressed in the whole germ cell bodies, but the protein migrates to the manchette, a unique structure at the base of elongating spermatid that directs formation of the flagella. In the elongating spermatids of wild-type mice, PACRG colocalizes with α-tubulin, a marker for the manchette, whereas this localization was not changed in the few remaining elongating spermatids of Meig1-deficient mice. In addition, MEIG1 no longer localizes to the manchette in the remaining elongating spermatids of Pacrg-deficient mice, indicating that PACRG recruits MEIG1 to the manchette. PACRG is not stable in mammalian cells, but can be stabilized by MEIG1 or by inhibition of proteasome function. SPAG16L is present in the spermatocyte cytoplasm of wild-type mice, and in the manchette of elongating spermatids, but in the Meig1 or Pacrg-deficient mice, SPAG16L no longer localizes to the manchette. By contrast, MEIG1 and PACRG are still present in the manchette of Spag16L-deficient mice, indicating that SPAG16L is a downstream partner of these two proteins. Together, our studies demonstrate that MEIG1/PACRG forms a complex in the manchette and that this complex is necessary to transport cargos, such as SPAG16L, to build the sperm flagella.
authors.affiliation: Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Otorhinolaryngology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Waixing
authors.initials: W
authors.lastname: Tang
authors.affiliation: Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Teves
authors.affiliation: Department of Infectious Diseases, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors.forename: Zhengang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei 430081, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Hongfei
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Biostatistics, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Kellie J
authors.initials: KJ
authors.lastname: Archer
authors.affiliation: Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Darrell L
authors.initials: DL
authors.lastname: Peterson
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Williams
authors.affiliation: Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298, USA Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298, USA.
authors.forename: Jerome F
authors.initials: JF
authors.lastname: Strauss
authors.affiliation: Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298, USA School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei 430081, China Department of Biochemistry & Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298, USA zzhang4@vcu.edu.
authors.forename: Zhibing
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D005407:Flagella / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D051379:Mice; D008817:Mice, Mutant Strains; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013091:Spermatogenesis / Q000235:genetics / Q000502:physiology; D020798:Two-Hybrid System Techniques
chemical_list: D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; C500770:Meig1 protein, mouse; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; C491467:PF20 protein, mouse; C471712:Pacrg protein, mouse; D010750:Phosphoproteins; D011506:Proteins
keywords: Cargo transport; Manchette; Sperm flagella; Spermiogenesis