Multiple protein kinase activities required for activation of sperm flagellar motility

Chaudhry, Prem S.; Creagh, Sue; Yu, Nam; Brokaw, Charles J.

doi:10.1002/cm.970320108

Cited by 29 publications

(26 citation statements)

References 32 publications

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“…A second group of Taf7l-regulated genes [Odf1, testis-specific serine kinase 6 (Tssk6), sperm motility kinase 2a/b (Smok2a/b), and Smok3a/b] have been implicated in sperm structure and motility during late stages of spermatogenesis ( Fig. 2 B and C) (27)(28)(29)(30). Additionally, a group of genes that have generally been classified as ones involved in metabolism are significantly down-regulated in the testis upon loss of Taf7l and likely reflect both metabolic and spermatogenesis functions.…”

Section: Resultsmentioning

confidence: 99%

Taf7l cooperates with Trf2 to regulate spermiogenesis

Zhou

Grubisic

Zheng

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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TATA-binding protein (TBP)-associated factor 7l (Taf7l; a paralogue of Taf7) and TBP-related factor 2 (Trf2) are components of the core promoter complex required for gene/tissue-specific transcription of protein-coding genes by RNA polymerase II. Previous studies reported that Taf7l knockout (KO) mice exhibit structurally abnormal sperm, reduced sperm count, weakened motility, and compromised fertility. Here we find that continued backcrossing of Taf7l −/Y mice from N5 to N9 produced KO males that are essentially sterile. Genome-wide expression profiling by mRNA-sequencing analysis of wild-type (WT) and Taf7l −/Y (KO) testes revealed that Taf7l ablation impairs the expression of many postmeiotic spermatogenic-specific as well as metabolic genes. Importantly, histological analysis of testes revealed that Taf7l −/Y mice develop postmeiotic arrest at the first stage of spermiogenesis, phenotypically similar to Trf2 −/− mice, but distinct from Taf4b −/− mice. Indeed, we find that Taf7l and Trf2 coregulate postmeiotic genes, but none of Taf4b-regulated germ stem cell genes in testes. Genome-wide ChIP-sequencing studies indicate that TAF7L binds to promoters of activated postmeiotic genes in testis. Moreover, biochemical studies show that TAF7L associates with TRF2 both in vitro and in testis, suggesting that TAF7L likely cooperates directly with TRF2 at promoters of a subset of postmeiotic genes to regulate spermiogenesis. Our findings thus provide a previously undescribed mechanism for cell-type-specific transcriptional control involving an interaction between a "nonprototypic" core promoter recognition factor (Trf2) and an orphan TAF subunit (Taf7l) in mammalian testis-specific gene transcription.gene regulation | reproduction | RNA-seq | contraceptive medicine S permatogenesis is a cyclic process in which diploid spermatogonia differentiate into mature haploid spermatozoa. This process is mainly driven by two-pre-and postmeiotictranscription waves that are tightly controlled by testis-specific transcription factors. During the premeiotic transcription phase, individual spermatogonia are committed to differentiating into primary spermatocytes that later undergo two meiotic divisions to generate haploid round spermatids connected by intercellular cytoplasmic bridges (1-3). During the postmeiotic transcription phase of spermiogenesis, haploid round spermatids are sculptured into the elongated shape of mature spermatozoa. These latter stages are accompanied by dramatic biochemical and morphological changes, including major remodeling of chromatin with protamines substituting for somatic histones to tightly pack DNA into the sperm nucleus.Understanding the intricate mechanisms that control spermatogenesis has important implications for human health and reproduction. A key step in the regulation of spermatogenesis occurs at the level of transcription, starting with the use of distinct promoter elements (4) within uniquely reorganized chromatin (5) and driven by the action of several testis-specific transcription factors...

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Section: Resultsmentioning

confidence: 99%

Taf7l cooperates with Trf2 to regulate spermiogenesis

Zhou

Grubisic

Zheng

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…Diverse experimental systems have revealed that ciliary and flagellar motility is controlled by phosphorylation and that the protein kinases and phosphatases responsible for regulation are anchored in the axoneme (27)(28)(29)(30)(31)(32)(33). Thus, we postulate that phosphorylation regulates dynein activity, and key kinases and phosphatases are anchored in the axoneme near the dynein arms or in the central pair/radial spoke structures.…”

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confidence: 88%

“…Therefore, we conclude that additional axonemal kinases are involved in the phosphorylation of IC138. Based on other studies (33,(35)(36)(37) and preliminary pharmacological analysis, we postulate one of the casein kinases (38) is anchored on doublet microtubules and directly phosphorylates IC138. In this study, we show that CKI is located in the axoneme, and inhibitors of CKI block phosphorylation of IC138 and rescue dynein activity in paralyzed axonemes.…”

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confidence: 88%

Casein Kinase I Is Anchored on Axonemal Doublet Microtubules and Regulates Flagellar Dynein Phosphorylation and Activity

Yang

Sale

2000

Journal of Biological Chemistry

105

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Flagellar dynein activity is regulated by phosphorylation. One critical phosphoprotein substrate in Chlamydomonas is the 138-kDa intermediate chain (IC138) of the inner arm dyneins (Habermacher, G., and Sale, W. S. (1997) J. Cell Biol. 136, 167-176). In this study, several approaches were used to determine that casein kinase I (CKI) is physically anchored in the flagellar axoneme and regulates IC138 phosphorylation and dynein activity. First, using a videomicroscopic motility assay, selective CKI inhibitors rescued dynein-driven microtubule sliding in axonemes isolated from paralyzed flagellar mutants lacking radial spokes. Rescue of dynein activity failed in axonemes isolated from these mutant cells lacking IC138. Second, CKI was unequivocally identified in salt extracts from isolated axonemes, whereas casein kinase II was excluded from the flagellar compartment. Third, Western blots indicate that within flagella, CKI is anchored exclusively to the axoneme. Analysis of multiple Chlamydomonas motility mutants suggests that the axonemal CKI is located on the outer doublet microtubules. Finally, CKI inhibitors that rescued dynein activity blocked phosphorylation of IC138. We propose that CKI is anchored on the outer doublet microtubules in position to regulate flagellar dynein.The dynein ATPase is a family of molecular motors responsible for diverse cellular functions including retrograde microtubule-based transport of organelles, assembly and function of the Golgi and mitotic apparatus, and movement of cilia and flagella (1, 2). One of the primary questions is how dynein activity is regulated. The focus of this report is based on a series of studies revealing that the flagellar central pair apparatus and radial spokes play a primary role in regulation of flagellar dynein activity (3, 4). The regulatory mechanism involves a structural network of axonemal kinases and phosphatases that control phosphorylation of key subunits within the inner arm dyneins (5-8). In particular, phosphorylation of a dynein intermediate chain subunit, IC138, 1 correlates with inhibition of flagellar dynein activity (7). This network of enzymes along with the central pair apparatus, radial spokes, and the inner arm dyneins operates to control flagellar waveform (9 -17). These conclusions are based on genetic analysis and in vitro functional studies of flagella from Chlamydomonas reinhardtii. For example, mutations that disrupt either the central pair or the radial spokes result in flagellar paralysis (2,3,18). These paralyzed axonemes undergo dynein-driven microtubule sliding in vitro (19) but at greatly reduced rates of sliding when compared with wild-type axonemes (20). Reconstitution and functional assays demonstrate that the radial spokes are required for wild-type dynein activity, and the velocity of dyneindriven microtubule sliding is mediated by posttranslational modification of the inner dynein arms (20). These results are consistent with other evidence indicating that the central pair/ radial spoke apparatus along with the dynein...

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“…Researchin several laboratories supportsthe ideathatreversibleproteinphosphorylation is associated with initiation andmodulationof sperm motility (1)(2)(3)(4)(5). Many studies aimed at examining the intracellular signaling pathways that initiate the activation of flagellar motility have relied on detergent-permeabilized sperm reactivated with exogenous 32p-ATP (2,6).…”

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confidence: 99%

“…However, the normal conditions of sperm collection and reactivation allow variable levels of motility to be expressed prior to analysis (7), as well as phosphorylation of many sperm proteins that are not clearly related to flagellar motility (1,8). Thus, identification of the few relevant proteins that are rapidly phosphorylated during the initial stages of activation is difficult.…”

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confidence: 99%