Maturation-promoting factor (abbreviated MPF, also called mitosis-promoting factor or M-Phase-promoting factor) is the cyclin-Cdk complex that was discovered first in frog eggs.
It stimulates the mitotic and meiotic phases of the cell cycle. MPF promotes the entrance into mitosis (the M phase) from the
G2 phase by phosphorylating multiple proteins needed during mitosis. MPF is activated at the end of G
2 by a
phosphatase
In biochemistry, a phosphatase is an enzyme that uses water to cleave a phosphoric acid Ester, monoester into a phosphate ion and an Alcohol (chemistry), alcohol. Because a phosphatase enzyme catalysis, catalyzes the hydrolysis of its Substrate ...
, which removes an inhibitory phosphate group added earlier.
The MPF is also called the M phase kinase because of its ability to phosphorylate target proteins at a specific point in the cell cycle and thus control their ability to function.
Discovery
In 1971, two independent teams of researchers (
Yoshio Masui and
Clement Markert
Clement Lawrence Markert (April 11, 1917 – October 1, 1999) was an American biologist credited with the discovery of isozymes (different forms of enzymes that catalyze the same reaction). He was a member of the National Academy of Sciences ...
, as well as Dennis Smith and Robert Ecker) found that frog
oocyte
An oocyte (, ), oöcyte, or ovocyte is a female gametocyte or germ cell involved in reproduction. In other words, it is an immature ovum, or egg cell. An oocyte is produced in a female fetus in the ovary during female gametogenesis. The female ...
s arrested in G
2 could be induced to enter M phase by microinjection of cytoplasm from oocytes that had been hormonally stimulated with progesterone.
Because the entry of oocytes into meiosis is frequently referred to as oocyte maturation, this cytoplasmic factor was called maturation promoting factor (MPF). Further studies showed, however, that the activity of MPF is not restricted to the entry of oocytes into meiosis. To the contrary, MPF is also present in somatic cells, where it induces entry into M phase of the mitotic cycle.
Evidence that a diffusible factor regulates the entry into mitosis had been previously obtained in 1966 using the slime mold ''Physarum polycephalum'' in which the nuclei of the multi-nucleate plasmodial form undergo synchronous mitoses. Fusing plasmodia whose cell cycles were out of phase with each other led to a synchronous mitosis in the next mitotic cycle. This result demonstrated that mitotic entry was controlled by a diffusible cytoplasmic factor and not by a "nuclear clock."
Structure
MPF is composed of two subunits:
*
Cyclin-dependent kinase 1
Cyclin-dependent kinase 1 also known as CDK1 or cell division cycle protein 2 homolog is a highly conserved protein that functions as a serine/threonine protein kinase, and is a key player in cell cycle regulation. It has been highly studied in th ...
(CDK1), the cyclin-dependent kinase subunit. It uses
ATP to phosphorylate specific
serine
Serine (symbol Ser or S) is an α-amino acid that is used in the biosynthesis of proteins. It contains an α-amino group (which is in the protonated − form under biological conditions), a carboxyl group (which is in the deprotonated − form un ...
and
threonine
Threonine (symbol Thr or T) is an amino acid that is used in the biosynthesis of proteins. It contains an α-amino group (which is in the protonated −NH form under biological conditions), a carboxyl group (which is in the deprotonated −COO ...
residues of target proteins.
*
Cyclin
Cyclin is a family of proteins that controls the progression of a cell through the cell cycle by activating cyclin-dependent kinase (CDK) enzymes or group of enzymes required for synthesis of cell cycle.
Etymology
Cyclins were originally disco ...
, a regulatory subunit. The cyclins are necessary for the kinase subunit to function with the appropriate substrate. The mitotic cyclins can be grouped as cyclins A & B. These cyclins have a nine residue sequence in the N-terminal region called the “destruction box”, which can be recognized by the ubiquitin ligase enzyme which destroys the cyclins when appropriate.
Role in the cell cycle
During G
1 and S phase, the CDK1 subunit of MPF is inactive due to an inhibitory enzyme, Wee1. Wee1 phosphorylates the Tyr-15 residues in yeast and Tyr-15 residues in humans of CDK1, rendering MPF inactive. During the transition of G
2 to M phase, cdk1 is de-phosphorylated by CDC25. The CDK1 subunit is now free and can bind to cyclin B, activate MPF, and make the cell enter mitosis. There is also a
positive feedback
Positive feedback (exacerbating feedback, self-reinforcing feedback) is a process that occurs in a feedback loop which exacerbates the effects of a small disturbance. That is, the effects of a perturbation on a system include an increase in the ...
loop that inactivates wee1.
Activation
MPF must be activated in order for the cell to transition from G
2 to M phase. There are three amino acid residues responsible for this G
2 to M phase transition. The Threonine-161 (Thr-161) on CDK1 must be phosphorylated by a
CDK-activating kinase
CDK-activating kinase (CAK) activates the cyclin- CDK complex by phosphorylating threonine residue 160 in the CDK activation loop. CAK itself is a member of the Cdk family and functions as a positive regulator of Cdk1, Cdk2, Cdk4, and Cdk6.
...
(CAK). CAK only phosphorylates Thr-161 when cyclin B is attached to CDK1.
In addition, two other residues on the CDK1 subunit must be activated by dephosphorylation. CDC25 removes a phosphate from residues Threonine-14 (Thr-14) and Tyrosine-15 (Tyr-15) and adds a hydroxyl group. Cyclin B/CDK1 activates CDC25 resulting in a positive feedback loop.
Overview of functions
* Triggers the formation of mitotic spindle through microtubule instability.
* Promotes mitosis i.e. chromatin condensation through phosphorylation of condensins.
* The three lamins present in the nuclear lamina, lamin A, B & C, are phosphorylated by MPF at serine amino residues. This leads to depolymerisation of the nuclear lamina & breakdown of nuclear envelope into small vesicles.
* Causes phosphorylation of GM130, which leads to the fragmentation of the Golgi and the ER.
Targets
The following are affected by MPF.
*
condensin
Condensins are large protein complexes that play a central role in chromosome assembly and segregation during mitosis and meiosis (Figure 1). Their subunits were originally identified as major components of mitotic chromosomes assembled in ''Xeno ...
s, which enable
chromatin
Chromatin is a complex of DNA and protein found in eukaryotic cells. The primary function is to package long DNA molecules into more compact, denser structures. This prevents the strands from becoming tangled and also plays important roles in r ...
condensation (see
prophase
Prophase () is the first stage of cell division in both mitosis and meiosis. Beginning after interphase, DNA has already been replicated when the cell enters prophase. The main occurrences in prophase are the condensation of the chromatin retic ...
)
* various
microtubule
Microtubules are polymers of tubulin that form part of the cytoskeleton and provide structure and shape to eukaryotic cells. Microtubules can be as long as 50 micrometres, as wide as 23 to 27 nm and have an inner diameter between 11 an ...
-associated proteins involved in
mitotic spindle
In cell biology, the spindle apparatus refers to the cytoskeletal structure of eukaryotic cells that forms during cell division to separate sister chromatids between daughter cells. It is referred to as the mitotic spindle during mitosis, a pr ...
formation
*
lamin
Lamins, also known as nuclear lamins are fibrous proteins in type V intermediate filaments, providing structural function and transcriptional regulation in the cell nucleus. Nuclear lamins interact with inner nuclear membrane proteins to form the ...
s, interaction contributing to degradation of the
nuclear envelope
The nuclear envelope, also known as the nuclear membrane, is made up of two lipid bilayer membranes that in eukaryotic cells surround the nucleus, which encloses the genetic material.
The nuclear envelope consists of two lipid bilayer membrane ...
* Histones, H
1 and H
3
* Golgi matrix, to cause fragmentation
Inhibition of myosin
MPF phosphorylates inhibitory sites on
myosin
Myosins () are a superfamily of motor proteins best known for their roles in muscle contraction and in a wide range of other motility processes in eukaryotes. They are ATP-dependent and responsible for actin-based motility.
The first myosin ...
early in mitosis. This prevents
cytokinesis
Cytokinesis () is the part of the cell division process during which the cytoplasm of a single eukaryotic cell divides into two daughter cells. Cytoplasmic division begins during or after the late stages of nuclear division in mitosis and meios ...
. When MPF activity falls at anaphase, the inhibitory sites are dephosphorylated and cytokinesis proceeds.
Disassembly by anaphase-promoting complex
MPF is disassembled when
anaphase-promoting complex
Anaphase-promoting complex (also called the cyclosome or APC/C) is an E3 ubiquitin ligase that marks target cell cycle proteins for degradation by the 26S proteasome. The APC/C is a large complex of 11–13 subunit proteins, including a culli ...
(APC) polyubiquitinates cyclin B, marking it for degradation in a negative feedback loop. In intact cells, cyclin degradation begins shortly after the onset of anaphase (late anaphase), the period of mitosis when sister chromatids are separated and pulled toward opposite spindle poles. As the concentration of Cyclin B/CDK1 increases, the heterodimer promotes APC to polyubiquitinate Cyclin B/CDK1.
References
{{Cell cycle proteins
Protein complexes
Cell cycle