The Drosophila melanogaster seminal fluid protease "seminase" regulates proteolytic and post-mating reproductive processes.
The Drosophila melanogaster seminal fluid protease "seminase" regulates proteolytic and post-mating reproductive processes.
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DOI：
10.1371/journal.pgen.1002435
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发表时间：
2012-01
期刊：
影响因子：
4.5
通讯作者：
Wolfner MF
中科院分区：
文献类型：
--
作者：
LaFlamme BA;Ram KR;Wolfner MF
从无脊椎动物到哺乳动物的各种动物的精液中都发现了蛋白水解酶及其抑制剂，它们是果蝇精液蛋白(SFP)中的主要蛋白质类。除了哺乳动物中调节精液凝块液化的单一蛋白酶级联外，在精液中还没有发现蛋白水解级联反应(即至少有两种蛋白水解酶作用的途径)。在果蝇中，SFP在交配过程中被转移到雌性体内，与精子一起，是雌性引发许多交配后反应所必需的。尽管几个SFP在交配过程中或交配后被蛋白分解，但几乎不知道参与这些切割事件的蛋白酶，也不知道精液中蛋白分解活动对雌性的生理后果。在这里，我们提出的证据表明，在交配期间和交配后，果蝇的精液中有一种蛋白水解酶的级联作用。使用RNAi下调SFP CG10586(一种预测的丝氨酸蛋白酶)的表达，我们发现它作用于SFP CG11864的上游，SFP CG11864是一种预测的ASTIN蛋白酶，处理与排卵和精子进入储藏有关的SFP。我们还表明，CG10586基因敲除会导致产卵水平降低，对后续雄性动物的性接受率增加，并导致精子使用模式异常，这些过程独立于CG11864。与CG10586基因敲除的雄性交配的雌性的长期表型与未能储存性肽的雌性相似，性肽是长期交配后反应的重要诱导者，这表明CG10586在调节性肽中发挥了作用。这些结果指出了昆虫SFP之间的蛋白分解的重要作用，并表明蛋白酶级联可能是一种精确的雌性交配后多个反应的时间调节机制。蛋白水解酶可以破坏、激活或以其他方式调节其他蛋白质的功能。在精液中，许多蛋白质在交配后必须被激活或降解；蛋白质分解是实现这一目标的有效方法，因为精液蛋白质在细胞外发挥作用，而大多数其他调节过程在细胞外无法使用。尽管许多动物的精液中都存在蛋白水解酶，但对它们所调控的过程几乎一无所知。在这里，我们提出了果蝇精液中的蛋白水解酶级联的证据。这一级联反应涉及两种在交配过程中被激活的蛋白酶。一旦在雌性体内，下游的蛋白酶就作用于另外两种蛋白质，这两种蛋白质对排卵和精子储存至关重要。有趣的是，级联顶部的蛋白酶CG10586也是雌性交配后的其他反应所必需的，包括产卵和精子使用，独立于第二种蛋白酶。因此，CG10586可能是雄性在交配后迅速激活许多雌性反应的通用调节开关。
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. Proteases can destroy, activate, or otherwise modulate the function of other proteins. In seminal fluid, many proteins have to be activated or degraded after mating; proteolysis is an effective way to accomplish this because seminal fluid proteins act outside of the cell, where most other regulatory processes cannot be used. Despite the presence of proteases in the seminal fluid of many animals, nearly nothing is known about the kinds of processes they regulate. Here, we present evidence of a protease cascade in the seminal fluid of the fruit fly Drosophila melanogaster. This cascade involves two proteases that are activated during mating. Once in the female, the downstream protease acts on two other proteins that are important for ovulation and sperm storage. Interestingly, the protease at the top of the cascade, CG10586, is also required for other female post-mating responses, including egg laying and sperm usage, independent of the second protease. Thus, CG10586 might be a general regulatory switch used by the male to quickly activate many female responses after mating.
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DOI：
10.1073/pnas.92.22.10114
发表时间：
1995-10-24
影响因子：
11.1
作者：
HERNDON, LA;WOLFNER, MF
通讯作者：
WOLFNER, MF
影响因子：
4.8
作者：
Guevara, Tibisay;Yiallouros, Irene;Xavier Gomis-Rueth, F.
通讯作者：
Xavier Gomis-Rueth, F.
影响因子：
3.3
作者：
Cui, Jun;Sackton, Katharine L.;Wolfner, Mariana F.
通讯作者：
Wolfner, Mariana F.
影响因子：
3.8
作者：
Kelleher, Erin S.;Watts, Thomas D.;Markow, Therese A.
通讯作者：
Markow, Therese A.
影响因子：
4.4
作者：
Dean MD;Findlay GD;Hoopmann MR;Wu CC;MacCoss MJ;Swanson WJ;Nachman MW
通讯作者：
Nachman MW